Substituted isoquinolinones

ABSTRACT

Isoquinolinone compounds are provided that are useful for the inhibition of ADP-platelet aggregation, particularly in the treatment of thrombosis and thrombosis related conditions or disorders.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of Provisional Application Ser. No.60/508,402, filed Oct. 3, 2003, the disclosure of which is incorporatedherein by reference.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH AND DEVELOPMENT

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REFERENCE TO A “SEQUENCE LISTING,” A TABLE, OR A COMPUTER PROGRAMLISTING APPENDIX SUBMITTED ON A COMPACT DISK

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BACKGROUND OF THE INVENTION

Thrombotic complications are a major cause of death in theindustrialized world. Examples of these complications include acutemyocardial infarction, unstable angina, chronic stable angina, transientischemic attacks, strokes, peripheral vascular disease,preeclampsia/eclampsia, deep venous thrombosis, embolism, disseminatedintravascular coagulation and thrombotic cytopenic purpura. Thromboticand restenotic complications also occur following invasive procedures,e.g., angioplasty, carotid endarterectomy, post CABG (coronary arterybypass graft) surgery, vascular graft surgery, stent placements andinsertion of endovascular devices and protheses. It is generally thoughtthat platelet aggregates play a critical role in these events. Bloodplatelets, which normally circulate freely in the vasculature, becomeactivated and aggregate to form a thrombus with disturbed blood flowcaused by ruptured atherosclerotic lesions or by invasive treatmentssuch as angioplasty, resulting in vascular occlusion. Plateletactivation can be initiated by a variety of agents, e.g., exposedsubendothelial matrix molecules such as collagen, or by thrombin whichis formed in the coagulation cascade.

An important mediator of platelet activation and aggregation is ADP(adenosine 5′-diphosphate) which is released from blood platelets in thevasculature upon activation by various agents, such as collagen andthrombin, and from damaged blood cells, endothelium or tissues.Activation by ADP results in the recruitment of more platelets andstabilization of existing platelet aggregates. Platelet ADP receptorsmediating aggregation are activated by ADP and some of its derivativesand antagonized by ATP (adenosine 5′-triphosphate) and some of itsderivatives (Mills, D. C. B. (1996) Thromb. Hemost. 76:835-856).Therefore, platelet ADP receptors are members of the family of P2receptors activated by purine and/or pyrimidine nucleotides (King, B.F., Townsend-Nicholson, A. & Burnstock, G. (1998) Trends Pharmacol. Sci.19:506-514).

Recent pharmacological data using selective antagonists suggests thatADP-dependent platelet aggregation requires activation of at least twoADP receptors (Kunapuli, S. P. (1998), Trends Pharmacol. Sci.19:391-394; Kunapuli, S. P. & Daniel, J. L. (1998) Biochem. J.336:513-523; Jantzen, H. M. et al. (1999) Thromb. Hemost. 81:111-117).One receptor appears to be identical to the cloned P2Y₁ receptor,mediates phospholipase C activation and intracellular calciummobilization and is required for platelet shape change. The secondplatelet ADP receptor important for aggregation mediates inhibition ofadenylyl cyclase. Molecular cloning of the gene or cDNA for thisreceptor (P2Y₁₂) has recently been reported (Hollopeter, G. et. al.(2001) Nature 409:202-207). Based on its pharmacological and signalingproperties this receptor has been previously termed P2Y_(ADP) (Fredholm,B. B. et al. (1997) TIPS 18:79-82), P2T_(AC) (Kunapuli, S. P. (1998),Trends Pharmacol. Sci. 19:391-394) or P2Ycyc (Hechler, B. et al. (1998)Blood 92, 152-159).

Various directly or indirectly acting synthetic inhibitors ofADP-dependent platelet aggregation with antithrombotic activity havebeen reported. The orally active antithrombotic thienopyridinesticlopidine and clopidogrel inhibit ADP-induced platelet aggregation,binding of radiolabeled ADP receptor agonist 2-methylthioadenosine5′-diphosphate to platelets, and other ADP-dependent events indirectly,probably via formation of an unstable and irreversible acting metabolite(Quinn, M. J. & Fitzgerald, D. J. (1999) Circulation 100: 1667-1667).Some purine derivatives of the endogenous antagonist ATP, e.g., AR-C(formerly FPL or ARL) 67085MX and AR-C69931MX, are selective plateletADP receptor antagonists which inhibit ADP-dependent plateletaggregation and are effective in animal thrombosis models (Humphries etal. (1995), Trends Pharmacol. Sci. 16, 179; Ingall, A. H. et al. (1999)J. Med. Chem. 42, 213-230). Novel triazolo[4,5-d]pyrimidine compoundshave been disclosed as P_(2T)—antagonists (WO 99/05144). Tricycliccompounds as platelet ADP receptor inhibitors have also been disclosedin WO 99/36425. The target of these antithrombotic compounds appears tobe the platelet ADP receptor mediating inhibition of adenylyl cyclase.

Despite these compounds, there exists a need for more effective plateletADP receptor inhibitors. In particular, there is a need for platelet ADPreceptor inhibitors having antithrombotic activity that are useful inthe prevention and/or treatment of cardiovascular diseases, particularlythose related to thrombosis.

BRIEF SUMMARY OF THE INVENTION

In view of the above, the present invention provides, in one aspect,compounds that are specifically substituted isoquinolinones. Thecompounds are represented by the formula:

With respect to formula I, the dotted line represents an optional doublebond; the symbol R¹ represents C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₅cycloalkyl, C₃₋₅ cycloalkyl-alkyl or benzyl; and the symbol R²represents H, C₁₋₆ alkyl or C₁₋₆ haloalkyl. The symbol R³ represents H,C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl, C₁₋₆ haloalkyl, C₁₋₆ hydroxyalkyl, cyano or—C(O)R^(3a), wherein R^(3a) is H, hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy,amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino; and R⁴ represents H orC₁₋₆ alkyl.

Turning next to the substituents on the thiophene ring, R⁵ represents H,halogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, cyano or —C(O)R^(5a), wherein R^(5a) represents C₁₋₆ alkoxy,amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino.

The symbol Ar represents an aromatic ring selected from benzene,pyridine and pyrimidine, each of which is optionally substituted withfrom 1-2 R⁶ substituents, wherein each R⁶ is independently selected fromhalogen, cyano, hydroxy, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆alkoxy, C₁₋₆ haloalkyl, C₁₋₆ haloalkoxy, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl, C₃₋₅ cycloalkyl-alkoxy, amino, C₁₋₆ alkylamino,di-C₁₋₆ alkylamino, —C(O)R^(6a), —O(CH₂)_(m)OR^(6b), —(CH₂)_(m)OR^(6b),—O(CH₂)_(m)N(R^(6b))₂ and —(CH₂)_(m)N(R^(6b))₂, wherein the subscript mis an integer of from 1 to 3, each R^(6a) is independently selected fromH, hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, amino, C₁₋₆ alkylamino and di-C₁₋₆alkylamino, and each R^(6b) is a member independently selected from thegroup consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl, and optionally, twoR^(6b) groups attached to nitrogen are combined with the nitrogen atomto form an azetidine, pyrrolidine or piperidine ring.

The present invention further provides pharmaceutically acceptable saltsof the above compounds, as well as pharmaceutical compositionscontaining those compounds.

In other aspects, the present invention provides methods of treatingthrombosis and thrombosis related conditions or disorders.

BRIEF DESCRIPTION OF THE DRAWINGS

Not applicable.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

The term “alkyl”, by itself or as part of another substituent, means,unless otherwise stated, a straight or branched chain hydrocarbonradical, having the number of carbon atoms designated (i.e. C₁₋₈ meansone to eight carbons). Examples of alkyl groups include methyl, ethyl,n-propyl, isopropyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl,n-hexyl, n-heptyl, n-octyl, and the like. The term “alkenyl” refers toan unsaturated alkyl group is one having one or more double bonds.Similarly, the term “alkynyl” refers to an unsaturated alkyl grouphaving one or more triple bonds. Examples of such unsaturated alkylgroups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl),2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl,3-butynyl, and the higher homologs and isomers. The term “cycloalkyl”refers to hydrocarbon rings having the indicated number of ring atoms(e.g., C₃₋₆cycloalkyl) and being fully saturated or having no more thanone double bond between ring vertices. When “cycloalkyl” is used incombination with “alkyl”, as in C₃₋₅ cycloalkyl-alkyl, the cycloalkylportion is meant to have from three to five carbon atoms, while thealkyl portion is an alkylene moiety having from one to three carbonatoms (e.g., —CH₂—, —CH₂CH₂— or —CH₂CH₂CH₂—).

The terms “alkoxy,” “alkylamino” and “alkylthio” (or thioalkoxy) areused in their conventional sense, and refer to those alkyl groupsattached to the remainder of the molecule via an oxygen atom, an aminogroup, or a sulfur atom, respectively. For brevity, the termC₁₋₆alkylamino is meant to include straight chain, branched or cyclicalkyl groups or combinations thereof, such as methyl, ethyl,2-methylpropyl, cyclobutyl and cyclopropylmethyl.

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” aremeant to include monohaloalkyl and polyhaloalkyl. For example, the term“C₁₋₄ haloalkyl” is mean to include trifluoromethyl,2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

The term “aryl” means, unless otherwise stated, a polyunsaturated,typically aromatic, hydrocarbon group which can be a single ring ormultiple rings (up to three rings) which are fused together or linkedcovalently. Exemplary aryl groups are phenyl, naphthyl, biphenyl and thelike. The term “heteroaryl” refers to aryl groups (or rings) thatcontain from one to five heteroatoms selected from N, O, and S, whereinthe nitrogen and sulfur atoms are optionally oxidized, and the nitrogenatom(s) are optionally quaternized. A heteroaryl group can be attachedto the remainder of the molecule through a heteroatom. Non-limitingexamples of heteroaryl groups include 1-pyrrolyl, 2-pyrrolyl,3-pyrrolyl, 1-pyrazolyl, 3-pyrazolyl, 2-imidazolyl, 4-imidazolyl,pyrazinyl, 2-oxazolyl, 4-oxazolyl, 5-oxazolyl, 3-isoxazolyl,4-isoxazolyl, 5-isoxazolyl, 2-thiazolyl, 4-thiazolyl, 5-thiazolyl,2-furyl, 3-furyl, 2-thienyl, 3-thienyl, 2-pyridyl, 3-pyridyl, 4-pyridyl,2-pyrimidyl, 4-pyrimidyl, 5-benzothiazolyl, purinyl, 2-benzimidazolyl,benzopyrazolyl, 5-indolyl, 1-isoquinolyl, 5-isoquinolyl, 2-quinoxalinyl,5-quinoxalinyl, 3-quinolyl, and 6-quinolyl. Substituents for each of theabove noted aryl and heteroaryl ring systems are selected from the groupof acceptable substituents described below.

As used herein, the term “heteroatom” is meant to include oxygen (O),nitrogen (N), sulfur (S) and silicon (Si).

The term “pharmaceutically acceptable salts” is meant to include saltsof the active compounds which are prepared with relatively nontoxicacids or bases, depending on the particular substituents found on thecompounds described herein. When compounds of the present inventioncontain relatively acidic functionalities, base addition salts can beobtained by contacting the neutral form of such compounds with asufficient amount of the desired base, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable base additionsalts include sodium, potassium, calcium, ammonium, organic amino, ormagnesium salt, or a similar salt. When compounds of the presentinvention contain relatively basic functionalities, acid addition saltscan be obtained by contacting the neutral form of such compounds with asufficient amount of the desired acid, either neat or in a suitableinert solvent. Examples of pharmaceutically acceptable acid additionsalts include those derived from inorganic acids like hydrochloric,hydrobromic, nitric, carbonic, monohydrogencarbonic, phosphoric,monohydrogenphosphoric, dihydrogenphosphoric, sulfuric,monohydrogensulfuric, hydriodic, or phosphorous acids and the like, aswell as the salts derived from relatively nontoxic organic acids likeacetic, propionic, isobutyric, malonic, benzoic, succinic, suberic,fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric,tartaric, methanesulfonic, and the like. Also included are salts ofamino acids such as arginate and the like, and salts of organic acidslike glucuronic or galactunoric acids and the like (see, for example,Berge, S. M., et al, “Pharmaceutical Salts”, Journal of PharmaceuticalScience, 1977, 66, 1-19). Certain specific compounds of the presentinvention contain both basic and acidic functionalities that allow thecompounds to be converted into either base or acid addition salts.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

In addition to salt forms, the present invention provides compoundswhich are in a prodrug form. Prodrugs of the compounds described hereinare those compounds that readily undergo chemical changes underphysiological conditions to provide the compounds of the presentinvention. Additionally, prodrugs can be converted to the compounds ofthe present invention by chemical or biochemical methods in an ex vivoenvironment. For example, prodrugs can be slowly converted to thecompounds of the present invention when placed in a transdermal patchreservoir with a suitable enzyme or chemical reagent.

Certain compounds of the present invention can exist in unsolvated formsas well as solvated forms, including hydrated forms. In general, thesolvated forms are equivalent to unsolvated forms and are intended to beencompassed within the scope of the present invention. Certain compoundsof the present invention may exist in multiple crystalline or amorphousforms. In general, all physical forms are equivalent for the usescontemplated by the present invention and are intended to be within thescope of the present invention.

Certain compounds of the present invention possess asymmetric carbonatoms (optical centers) or double bonds; the racemates, diastereomers,geometric isomers and individual isomers (e.g., separate enantiomers)are all intended to be encompassed within the scope of the presentinvention.

The compounds of the present invention may also contain unnaturalproportions of atomic isotopes at one or more of the atoms thatconstitute such compounds. For example, the compounds may beradiolabeled with radioactive isotopes, such as for example tritium(³H), iodine-125 (¹²⁵I) or carbon-14 (¹⁴C). All isotopic variations ofthe compounds of the present invention, whether radioactive or not, areintended to be encompassed within the scope of the present invention.

General

DESCRIPTION OF THE EMBODIMENTS

Compounds

In view of the above, the present invention provides, in one aspect,compounds that are specifically substituted isoquinolinones. Thecompounds are represented by the formula:

Turning first to the symbols R¹ through R⁴, the symbol R¹ representsC₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl orbenzyl. For the terms C₃₋₅ cycloalkyl-alkyl and (as used below) C₃₋₅cycloalkyl-alkoxy, the alkyl or alkoxy portions respectively are meantto have from one to three carbon atoms, exclusive of the carbon atomsused in the cycloalkyl portion. For example, C₃₋₅ cycloalkyl-alkyl ismeant to include cyclopropylmethyl, cyclopentylmethyl,3-cyclobutylpropyl, 2-cylcopropylethyl, and the like. Similarly, C₃₋₅cycloalkyl-alkoxy is meant to include cyclopropylmethoxy,cyclopentylmethoxy, 3-cyclobutylpropyloxy, 2-cylcopropylethoxy, and thelike. Preferably, R¹ is C₁₋₆ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl. More preferably, R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, orC₃₋₅ cycloalkyl-alkyl. Still more preferably, R¹ is C₁₋₄ alkyl,particularly CH₃ or CH₂CH₃ with CH₃ being the most preferred.

The symbol R² represents H, C₁₋₆ alkyl or C₁₋₆ haloalkyl. Preferably, R²is H or C₁₋₆ alkyl; more preferably H or C₁₋₄ alkyl. Still morepreferably, R² is H or CH₃, with H being the most preferred.

The symbol R³ represents H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₅cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₆ haloalkyl, C₁₋₆ hydroxyalkyl,cyano or —C(O)R^(3a), wherein R^(3a) is selected from H, hydroxy, C₁₋₆alkyl, C₁₋₆ alkoxy, amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino.Preferably, R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyano or —C(O)R^(3a).More preferably, R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅cycloalkyl, or C₃₋₅ cycloalkyl-alkyl.

The symbol R⁴ represents H or C₁₋₆ alkyl. Preferably, R⁴ is H or C₁₋₄alkyl. More preferably, R⁴ is H or CH₃.

Turning next to the substituents on the thiophene ring, R⁵ represents H,halogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ haloalkyl, C₁₋₆alkoxy, cyano or —C(O)^(Ra), wherein R^(5a) represents C₁₋₆ alkoxy,amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino. Preferably, R⁵ is H,halogen, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ haloalkyl, C₁₋₄alkoxy, cyano or —C(O)R^(5a). More preferably, R⁵ is halogen, C₁₋₄alkyl, C₁₋₄ haloalkyl, C₁₋₄ alkoxy, cyano, —C≡CH or —C(O)NH₂. Still morepreferably, R⁵ is halogen or C₁₋₄ alkyl. Most preferably, R⁵ is chloro,and is attached to the 5-position of the thienyl ring.

The symbol Ar represents an aromatic ring selected from benzene,pyridine and pyrimidine, each of which is optionally substituted withfrom 1-2 R⁶ substituents, wherein each R⁶ is independently selected fromhalogen, cyano, hydroxy, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆alkoxy, C₁₋₆ haloalkyl, C₁₋₆ haloalkoxy, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl, C₃₋₅ cycloalkyl-alkoxy, amino, C₁₋₆ alkylamino,di-C₁₋₆ alkylamino, —C(O)R^(6a), —O(CH₂)_(m)OR^(6b), —(CH₂)_(m)OR^(6b),—O(CH₂)_(m)N(R^(6b))₂ and —(CH₂)_(m)N(R⁶)₂, wherein the subscript m isan integer of from 1 to 3, each R^(6a) is independently selected from H,hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, amino, C₁₋₆ alkylamino and di-C₁₋₆alkylamino, and each R^(6b) is independently selected from H, C₁₋₄ alkyland C₁₋₄ alkanoyl, and optionally, two R^(6b) groups attached tonitrogen are combined with the nitrogen atom to form an azetidine,pyrrolidine or piperidine ring. Each of the aromatic rings (optionallysubstituted) is a separate and preferred embodiment of the presentinvention.

The dotted line in Formula I represents an optional double bond. In mostembodiments, the double bond is present, and preferred. In someembodiments, however, the double bond is not present, the remainingvalences being filled with hydrogen atoms. As a result, the dotted lineis meant to represent both of the following:

A number of particularly preferred embodiments are provided as formulaeIa, Ib and Ic.

In a first group of preferred embodiments, the compounds of the presentinvention have the formula:

wherein each of R¹, R², R³, R⁴, R⁵, and R⁶ have the meanings providedabove, and the subscript n is an integer of from 0 to 2, indicating theabsence (n is 0) or presence (n is 1 or 2) of substituents that areindependently selected from the groups provided above for R⁶. Furtherpreferred are those embodiments in which R⁶, when present, occupypositions on the benzene ring that are adjacent to the carbon atombearing the urea-sulfonyl (—NHC(O)NHS(O)₂—) component. Additionally,preferred components provided above with respect to the general formulaI are also preferred for compounds of formula Ia.

In one group of preferred embodiments of formula Ia, n is an integer offrom 0 to 2; R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoor —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkyl-alkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ whereinthe subscript m is 1 or 2 and each R^(6b) is independently selected fromthe group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl. Still furtherpreferred are those embodiments in which R¹ is C₁₋₄ alkyl; R⁴ is H orCH₃; R⁵ is halogen or C₁₋₄ alkyl; and each R⁶ when present is selectedfrom C₁₋₄ alkyl, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂. Evenfurther preferred are those embodiments in which R¹ is methyl; R⁵ ischloro, and is attached at the 5-position of the thienyl ring; and eachR⁶ when present is selected from CH₃, —OCH₂CH₂OH, —OCH₂CH₂OCH₃,—OCH₂OCH₃, —OCH₂CH₂OC(O)CH₃ and —O(CH₂)₂N(CH₃)₂. In separate, butpreferred groups of embodiments, the subscript n is 0, or 1, or 2.

In a second group of preferred embodiments, the compounds of the presentinvention have the formula:

wherein each of R¹, R², R³, R⁴, R⁵, and R⁶ have the meanings providedabove, and the subscript n is an integer of from 0 to 2, indicating theabsence (n is 0) or presence (n is 1 or 2) of substituents that areindependently selected from the groups provided above for R⁶. Furtherpreferred are those embodiments in which R⁶, when present as a singlesubstituent, occupies the 3-position on the pyridine ring (i.e., thatposition adjacent to the carbon atom bearing the urea-sulfonyl(—NHC(O)NHS(O)₂—) component). Additionally, preferred componentsprovided above with respect to the general formula I are also preferredfor compounds of formula Ib.

In one group of preferred embodiments of formula Ib, n is an integer offrom 0 to 2; R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoor —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkyl-alkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R⁶)₂ wherein thesubscript m is 1 or 2 and each R^(6b) is independently selected from thegroup consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl. Still furtherpreferred are those embodiments in which R¹ is C₁₋₄ alkyl; R⁴ is H orCH₃; R⁵ is halogen or C₁₋₄ alkyl; and each R⁶ when present is selectedfrom C₁₋₄ alkyl, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂. Evenfurther preferred are those embodiments in which R¹ is methyl; R³ is H,C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl or C₃₋₅cycloalkyl-alkyl; R⁴ is H or CH₃; R⁵ is chloro and is attached at the5-position of the thienyl ring; and R⁶, when present is selected fromC₁₋₄ alkyl, O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ wherein thesubscript m is 1 or 2 and each R^(6b) is independently selected from H,C₁₋₄ alkyl and C₁₋₄ alkanoyl.

In still other embodiments, the compounds of the invention have theformula Ic:

wherein each of R¹, R², R³, R⁴, R⁵, and R⁶ have the meanings providedabove, and the subscript n is an integer of from 0 to 2, indicating theabsence (n is 0) or presence (n is 1 or 2) of substituents that areindependently selected from the groups provided above for R⁶. Preferredcomponents provided above with respect to the general formula I are alsopreferred for compounds of formula Ic.

In one group of preferred embodiments of formula Ic, n is an integer offrom 0 to 2; R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoor —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkyl-alkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ whereinthe subscript m is 1 or 2 and each R^(6b) is independently selected fromthe group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl. Still furtherpreferred are those embodiments in which R¹ is C₁₋₄ alkyl; R⁴ is H orCH₃; R⁵ is halogen or C₁₋₄ alkyl; and each R⁶ when present is selectedfrom C₁₋₄ alkyl, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂. Evenfurther preferred are those embodiments in which R¹ is methyl; R³ is H,CIA alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl or C₃₋₅cycloalkyl-alkyl; R⁴ is H or CH₃; R⁵ is chloro and is attached at the5-position of the thienyl ring; and R⁶, when present is selected from C₄alkyl, O(CH₂)_(m)OR^(6b) and O(CH₂)_(m)N(R^(6b))₂ wherein the subscriptm is 1 or 2 and each R^(6b) is independently selected from H, C₁₋₄ alkyland C₁₋₄ alkanoyl.

Among the most preferred embodiments of the invention are the compoundsprovided below, as well as in the Examples.

Scheme A describes a method of preparing compounds of Formula I whereinR⁴═H, R¹, R², R³, R⁵ is described hereinbefore, Ar is substituted aryland heteroaryl.

A compound of Formula I can be prepared by reacting malonic acid andbenzaldehyde 1 in pyridine to provide cinnamic acid 2, which can beconverted to an acryloyl azide by first treating with ethylchloroformate then sodium azide. Curtius rearrangement and cyclizationof acryloyl azide, in the presence of a catalyst such as iodine in aninert solvent such as 1,2-dichlorobenzene provides isoquinolone 3. Thesubstituted isoquinolone 4 can be prepared by treating the amidefunctionality of isquinolone 3 with a halogen substituted aromatic orheteroaromatic compound, wherein the halogen is a leaving group,preferably chloro or fluoro, in the presence of a base such as potassiumcarbonate or cesium carbonate. Preferred solvents for thistransformation are inert solvents such as DMF, DMSO, and lower alcohols.The methyl group can then be removed by treatment of BBr₃ indichloromethane, or alternatively lithium iodide in an inert solventsuch as DMSO or DMF. The C—N coupling reaction of the aryltriflate 5with carbamic acid tert-butyl ester, or primary or secondary amines canbe carried out according to methods described in Buchwald et al., Org.Lett. 2000, 2, 1101-1104. The products from coupling reaction of thearyltriflate 5 with carbamic acid tert-butyl ester can be alkylatedunder basic condition. The nitro group of compound 6 can be reduced byprocedures known to one skilled in the art to yield a free amino group.For example, one suitable method of reduction involves hydrogenation,with a suitable catalyst (e.g., 10% palladium on carbon) in anappropriate solvent, typically an alcohol. The formation of thesulfonylurea linkage can be accomplished by treating the reduced productaniline 7 with a pre-mixed solution of 5-chlorothiophene-2-sulfonamide,N,N′-disuccinimidyl carbonate and tetramethylguanidine indichloromethane, followed by treatment with TFA in dichloromethane atroom temperature to afford the sulfonylurea 8.

A compound of Formula I with varying Ar groups can be prepared by firstsynthesizing the common intermediate 11 in 6 steps (see Scheme B).Compound 3 from Scheme A can be demethylated by treatment with borontribromide in dichloromethane, followed by selective triflation withphenyltrifluoromethylsulfonimide to give the triflate 9. Protection ofthe lactam nitrogen, with SEM-Cl and C—N coupling using carbamic acidtert-butyl ester can be carried out according to Buchwald et al., Org.Lett. 2000, 2, 1101-1104, to give bis-protected intermediate 10.Standard methylating conditions and removal of the SEM group with TBAFprovides the key intermediate 11. A variety of halo-substitutednitroaromatic compounds can be coupled with 11 using Method A or Bconditions, followed by reduction using catalytic hydrogenation ortin(II) dichloride dihydrate to give 12 (see, Examples 46 and 47 below).Also, a variety of halo-substituted anilines can be coupled to 10 usingMethod C conditions as outlined below to give 12 (see, Example 48). Theformation of sulfonylurea linkage can be accomplished by treating theproduct aniline 12 with the ethyl carbamate of5-chloro-thiophene-2-sulfonamide in refluxing toluene, followed bytreatment with TFA in dichloromethane at room temperature to afford thesulfonylurea 13.

Scheme C describes a method of preparing a compounds of Formula Iwherein R⁴═H, R¹═Me, R²═H, R⁵═Cl, R³ is described hereinbefore. Ar is asubstituted or unsubstituted aryl or heteroaryl group.

As seen in Scheme C, compounds of Formula I can be prepared by startingwith deprotection of the t-Boc group of compound 14 which can be readilyobtained from Scheme B, followed by halogenation to provide compound 15.Conversion of 15 to compound 16 can be accomplished using Stille orSuzuki coupling conditions to provide compound 16 with appropriatelysubstituted R³ group. The formation of a sulfonylurea linkage can beaccomplished by treating the reduced product aniline 7 with a pre-mixedsolution of 5-chlorothiophene-2-sulfonamide, N,N′-disuccinimidylcarbonate and tetramethylguanidine in dichloromethane, followed bytreatment with TFA in dichloromethane at room temperature to afford thesulfonylurea 17.

Compositions

In another aspect of the invention, pharmaceutical compositions areprovided in which compounds of formulae I, Ia, Ib, or Ic, alone or incombination, are combined with a pharmaceutically acceptable carrier.Preferred compounds for use in the compositions of the present inventionare those compounds identified above as specific or preferredembodiments.

The pharmaceutical compositions of the invention may be in the form ofsolutions or suspensions. In the management of thrombotic disorders thecompounds or pharmaceutical compositions of the invention may also be insuch forms as, for example, tablets, capsules or elixirs for oraladministration, suppositories, sterile solutions or suspensions orinjectable administration, and the like, or incorporated into shapedarticles.

Typical adjuvants which may be incorporated into tablets, capsules andthe like include, but are not limited to, binders such as acacia, cornstarch or gelatin, and excipients such as microcrystalline cellulose,disintegrating agents like corn starch or alginic acid, lubricants suchas magnesium stearate, sweetening agents such as sucrose or lactose, orflavoring agents. When a dosage form is a capsule, in addition to theabove materials it may also contain liquid carriers such as water,saline, or a fatty oil. Other materials of various types may be used ascoatings or as modifiers of the physical form of the dosage unit.Sterile compositions for injection can be formulated according toconventional pharmaceutical practice. For example, dissolution orsuspension of the active compound in a vehicle such as an oil or asynthetic fatty vehicle like ethyl oleate, or into a liposome may bedesired. Buffers, preservatives, antioxidants and the like can beincorporated according to accepted pharmaceutical practice.

Additionally, dosage formulations of compounds of formulae I, Ia, Ib, orIc, or pharmaceutical compositions containing a compound of theinvention, to be used for therapeutic administration must be sterile.Sterility can be readily accomplished by filtration through sterilemembranes such as 0.2 micron membranes, or by other conventionalmethods. Formulations typically will be stored in a solid form,preferably in a lyophilized form. While the preferred route ofadministration is orally, the dosage formulations of compounds offormulae I, Ia, Ib, or Ic, or pharmaceutical compositions of theinvention may also be administered by injection, intravenously (bolusand/or infusion), subcutaneously, intramuscularly, colonically,rectally, nasally, transdermally or intraperitoneally. A variety ofdosage forms may be employed as well including, but not limited to,suppositories, implanted pellets or small cylinders, aerosols, oraldosage formulations and topical formulations such as ointments, dropsand dermal patches. The compounds of formulae I, Ia, Ib, or Ic, andpharmaceutical compositions of the invention may also be incorporatedinto shapes and articles such as implants which may employ inertmaterials such biodegradable polymers or synthetic silicones as, forexample, SILASTIC, silicone rubber or other polymers commerciallyavailable. The compounds and pharmaceutical compositions of theinvention may also be provided in the form of liposome delivery systems,such as small unilamellar vesicles, large unilamellar vesicles andmultilamellar vesicles. Liposomes can be formed from a variety oflipids, such as cholesterol, stearylamine or phosphatidylcholines, usedmethods well known to one of skill in the art.

Methods of Treatment/Administration

In yet another aspect, the present invention provides methods forpreventing or treating thrombosis in a mammal by administering to themammal a therapeutically effective amount of a compound of formulae I,Ia, Ib, or Ic, alone or as part of a pharmaceutical composition of theinvention as described above. Compounds of formulae I, Ia, Ib, or Ic,and pharmaceutical compositions of the invention containing a compoundof formulae I, Ia, Ib, or Ic, of the invention are suitable for usealone or as part of a multi-component treatment regimen for theprevention or treatment of cardiovascular diseases, particularly thoserelated to thrombosis. For example, a compound or pharmaceuticalcomposition of the invention may be used as a drug or therapeutic agentfor any thrombosis, particularly a platelet-dependent thromboticindication, including, but not limited to, acute myocardial infarction,unstable angina, chronic stable angina, transient ischemic attacks,strokes, peripheral vascular disease, preeclampsia/eclampsia, deepvenous thrombosis, embolism, disseminated intravascular coagulation andthrombotic cytopenic purpura, thrombotic and restenotic complicationsfollowing invasive procedures, e.g., angioplasty, carotidendarterectomy, post CABG (coronary artery bypass graft) surgery,vascular graft surgery, stent placements and insertion of endovasculardevices and protheses.

Compounds and pharmaceutical compositions of the invention may also beused as part of a multi-component treatment regimen in combination withother therapeutic or diagnostic agents in the prevention or treatment ofthrombosis in a mammal. In certain preferred embodiments, compounds orpharmaceutical compositions of the invention may be coadministered alongwith other compounds typically prescribed for these conditions accordingto generally accepted medical practice such as anticoagulant agents,thrombolytic agents, or other antithrombotics, including plateletaggregation inhibitors, tissue plasminogen activators, urokinase,prourokinase, streptokinase, heparin, aspirin, or warfarin. Still otheragents that can be administered with the compounds of the presentinvention include antiplatelet compounds, fibrinolytics,anti-inflammatory compounds, cholesterol-lowering agents,blood-pressure-lowering agents and serotonin blockers. Suitableantiplatelet compounds include GPIIB-IIIa antagonists, aspirin,phosphodiesterase III inhibitors and thromboxane A2 receptorantagonists. Suitable anticoagulants include thrombin inhibitors,coumadin (Warfarin), heparin and Lovenox®. Suitable anti-inflammatorycompounds include non-steroidal anti-inflammatory agents,cyclooxygenase-2 inhibitors and rheumatoid arthritis agents.Coadministrations of these agents with the compounds of the inventionmay also allow for application of reduced doses of the thrombolyticagents and therefore minimize potential hemorrhagic side-effects.Compounds and pharmaceutical compositions of the invention may also actin a synergistic fashion to prevent reocclusion following a successfulthrombolytic therapy and/or reduce the time to reperfusion.

In related methods, the compounds of the invention are useful for theprevention of a secondary ischemic event. In these methods, compounds ofthe invention or their pharmaceutical compositions are administered to apatient who has suffered a primary ischemic event in an amountsufficient to prevent or reduce the likely occurrence of a secondaryevent. Generally, the primary and/or secondary ischemic event isselected from myocardial infraction, stable or unstable angina, acutereocclusion after percutaneous transluminal coronary angioplasty,restenosis, thrombotic stroke, transient ischemic attack, reversibleischemic neurological deficit and intermittent claudication.

The compounds and pharmaceutical compositions of the invention may beutilized in vivo, ordinarily in mammals such as primates, (e.g.,humans), sheep, horses, cattle, pigs, dogs, cats, rats and mice, or invitro. The biological properties, as defined above, of a compound or apharmaceutical composition of the invention can be readily characterizedby methods that are well known in the art such as, for example, by invivo studies to evaluate antithrombotic efficacy, and effects onhemostasis and hematological parameters.

Subjects (typically mammalian) in need of treatment using the compoundsor pharmaceutical compositions of the invention may be administereddosages that will provide optimal efficacy. The dose and method ofadministration will vary from subject to subject and be dependent uponsuch factors as the type of mammal being treated, its sex, weight, diet,concurrent medication, overall clinical condition, the particularcompound of formulae I, Ia, Ib, or Ic employed, the specific use forwhich the compound or pharmaceutical composition is employed, and otherfactors which those skilled in the medical arts will recognize.

Therapeutically effective dosages may be determined by either in vitroor in vivo methods. For each particular compound or pharmaceuticalcomposition of the invention, individual determinations may be made todetermine the optimal dosage required. The range of therapeuticallyeffective dosages will be influenced by the route of administration, thetherapeutic objectives and the condition of the patient. For injectionby hypodermic needle, it may be assumed the dosage is delivered into thebodily fluids. For other routes of administration, the absorptionefficiency must be individually determined for each compound by methodswell known in pharmacology. Accordingly, it may be necessary for thetherapist to titer the dosage and modify the route of administration asrequired to obtain the optimal therapeutic effect.

The determination of effective dosage levels, that is, the dosage levelsnecessary to achieve the desired result, i.e., platelet ADP receptorinhibition, will be readily determined by one skilled in the art.Typically, applications of a compound or pharmaceutical composition ofthe invention are commenced at lower dosage levels, with dosage levelsbeing increased until the desired effect is achieved. The compounds andcompositions of the invention may be administered orally in an effectiveamount within the dosage range of about 0.01 to 1000 mg/kg in a regimenof single or several divided daily doses. If a pharmaceuticallyacceptable carrier is used in a pharmaceutical composition of theinvention, typically, about 5 to 500 mg of a compound of formulae I, Ia,Ib, or Ic, is compounded with a pharmaceutically acceptable carrier ascalled for by accepted pharmaceutical practice including, but notlimited to, a physiologically acceptable vehicle, carrier, excipient,binder, preservative, stabilizer, dye, flavor, etc. The amount of activeingredient in these compositions is such that a suitable dosage in therange indicated is obtained.

The following preparations and examples are given to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representativethereof.

EXAMPLES

The starting materials and reagents used in preparing these compoundsgenerally are either available from commercial suppliers, such asAldrich Chemical Co., or are prepared by methods known to those skilledin the art following procedures set forth in references such as Fieserand Fieser 's Reagents for Organic Synthesis; Wiley & Sons: New York,1991, Volumes 1-15; Rodd's Chemistry of Carbon Compounds, ElsevierScience Publishers, 1989, Volumes 1-5 and Supplementals; and OrganicReactions, Wiley & Sons: New York, 1991, Volumes 1-40. The followingsynthetic reaction schemes are merely illustrative of some methods bywhich the compounds of the present invention can be synthesized, andvarious modifications to these synthetic reaction schemes can be madeand will be suggested to one skilled in the art having referred to thedisclosure contained in this Application.

The starting materials and the intermediates of the synthetic reactionschemes can be isolated and purified if desired using conventionaltechniques, including but not limited to, filtration, distillation,crystallization, chromatography, and the like. Such materials can becharacterized using conventional means, including physical constants andspectral data.

Unless specified to the contrary, the reactions described hereinpreferably are conducted under an inert atmosphere at atmosphericpressure at a reaction temperature range of from about −78° C. to about150° C., more preferably from about 0° C. to about 125° C., and mostpreferably and conveniently at about room (or ambient) temperature,e.g., about 20° C.

Example 1 3-(4-Fluoro-3-methoxyphenyl)-acrylic Acid

To a solution of 4-fluoro-3-methoxybenzaldehyde (32 g, 0.2 mol) inpyridine (100 mL) was added malonic acid (43 g, 0.4 mol) and piperidine(3 mL, 0.03 mol). The reaction solution was stirred at 85° C. for 13 hr.Upon cooling, the resulting suspension was added to cold water (500 mL)and acidified with conc. HCl (80 mL). The white solid was filtered off,washed with water and dried to yield 36 g (92%) of3-(4-fluoro-3-methoxyphenyl)-acrylic acid. RP-HPLC: 3.71 min. 1H-NMR(DMSO-d₆) δ (ppm) 3.85 (s, 3), 6.53 (d, 1, J=16), 7.20 (m, 2), 7.50 (m,1), 7.52 (d, 1, J=16).

Example 2 3-(4-Fluoro-3-methoxyphenyl)-acryloyl Azide

To a chilled solution (ice/acetone) of3-(4-fluoro-3-methoxyphenyl)-acrylic acid (37 g, 0.194 mmol) in dry THF(280 mL) and triethylamine (352 mL) was added ethyl chloroformate (22.5mL) in THF (50 mL) dropwise over 20 minutes. The resulting suspensionwas allowed to warm to 23° C. for 1 h, re-cooled, and a solution of NaN₃(18.7 g) in water (80 mL) was added. The reaction was stirred at 23° C.for 1-2 hr. Workup involved addition of dichloromethane (250 mL)followed by incremental slow addition of 1N HCl. The aqueous layer wasfurther extracted 2 times with dichloromethane. The organic layers werecombined, washed with 1N HCl and brine, dried over MgSO₄, andconcentrated in vacuo to afford 39.8 g (95%) of the acyl azide. RP-HPLC:5.31 min.

Example 3 7-Fluoro-6-methoxy-2H-isoquinolin-1-one

A solution of the acyl azide (39 g, see Example 2) in1,2-dichlorobenzene (300 mL) was heated to 140° C. for approximately 1 huntil gas formation subsides. Catalytic iodine was added and thetemperature was increased to 180° C. for 1.5 h. The reaction mixture wasallowed to cool to ambient temperature with stirring; the precipitatewhich formed was collected by filtration, washed with benzene and driedunder vacumn to afford 22.6 g (67%) of7-fluoro-6-methoxy-2H-isoquinolin-1-one as a tan solid. RP-HPLC: 2.58min; ES-MS (M+H)⁺=194.1; 1H-NMR (DMSO-d₆) δ(ppm): 3.8 (3H, s), 6.48(1,d), 7.11 (t, 1), 7.33 (d, 1); 6.77 (d, 1).

Example 4 7-Fluoro-6-methoxy-2-(4-nitrophenyl)-2H-isoquinolin-1-one

To a solution of 11.0 g of 7-Fluoro-6-methoxy-2H-isoquinolin-1-one (57mmol) in DMF (108 mL) was added potassium carbonate (11.8 g), followedby 10.6 g of 1-fluoro-4-nitrobenzene (75 mmol). The reaction mixture wasstirred at 120° C. for 6 hr then poured onto ice water. The slurry wasextracted with ether to remove excess pFPhNO₂. The precipitate wascollected by filtration, washed with ether and dried in vacuo to give12.1 g (68%) of the product asa yellow solid. RP-HPLC: 4.79 min; ES-MS(M+H)⁺=315.0; 1H-NMR (DMSO-d₆) δ (ppm): 4.0 (3H, s), 6.76 (1,d), 7.48(d, 1), 7.53 (d, 1), 7.83 (d,2), 7.92 (d, 1), 8.38 (d, 2).

Example 5 7-Fluoro-6-hydroxy-2-(4-nitro-phenyl)-2H-isoquinolin-1-one

To a chilled suspension of7-Fluoro-6-methoxy-2-(4-nitrophenyl)-2H-isoquinolin-1-one (3.14 g, 10mmol) in dichloromethane (50 mL) was added neat boron tribromide (8 mL,85 mmol) via syringe. The brown suspension was stirred at roomtemperature for 24 hr. The solvent was decanted, washed with cold DCM,leaving a black residue, which was triturated on ice with methanol (80mL). The solid was collected by filtration, then washed with water anddried to give 2.66 g (89%) of7-Fluoro-6-hydroxy-2-(4-nitro-phenyl)-2H-isoquinolin-1-one. RP-HPLC:3.93 min; ES-MS (M+H)⁺=301.0; 1H-NMR (DMSO-d₆) δ (ppm): 6.66 (d,1), 7.16(d, 1), 7.43 (d, 1); 7.76 (d, 2), 7.84 (d, 1), 8.33 (d, 2), 11.1 (br s,1).

Example 6 Trifluoro-methanesulfonic Acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl Ester

To a suspension of7-Fluoro-6-hydroxy-2-(4-nitro-phenyl)-2H-isoquinolin-1-one (1.15 g, 3.8mmol) in dry pyridine (25 mL) and dichloromethane (20 mL) was added neattrifluoromethanesulfonic anhydride (0.8 mL, 4.76 mmol) dropwise over 5min. The resulting solution was stirred at room temp for 2 hr. Thereaction mixture was diluted with ethyl acetate (200 mL), washed with 1NHCl (60 mL), water (50 mL) and brine (50 mL), dried over sodium sulfate,filtered, concentrated in vacuo and dried to give 1.37 g (83%) of puretrifluoromethanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester.RP-HPLC: 6.20 min; ES-MS (M+H)⁺=433.0; 1H-NMR (DMSO-d₆) δ (ppm): 6.88(d, 1), 7.64 (d, 1), 7.80 (d, 2), 8.24 (m, 2, 8.37 (d, 2).

Example 7[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicAcid Tert-Butyl Ester

In a dry flask was combined trifluoromethanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(0.86 g, 2 mmol), t-butyl carbamate (0.33 g, 2.8 mmol), dry powderedcesium carbonate (1.1 g, 3.4 mmol),9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (Xantphos, 0.14 g, 0.24mmol), and tris(dibenzylideneacetone)dipalladium(0) (Pd₂ dba₃, 38 mg,0.08 mmol). Under Ar atmosphere, dry THF (17 mL) was added to the flask,and the mixture was stirred at 75° C. for 25 hr. The reaction wasconcentrated and purified on silica gel using EtOAc/hexane as eluent togive 0.64 g (80%) of pure[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester as a white solid. RP-HPLC: 5.98 min; ES-MS(M+H)⁺=400.0; 1H-NMR (DMSO-d₆) δ (ppm): 1.47 (s, 9), 6.77 (d, 1), 7.46(d, 1), 7.78 (d, 2), 7.87 (d, 1), 8.17 (d, 1), 8.33 (d, 2), 9.54 (s, 1).

Example 8[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicAcid Tert-Butyl Ester

To a solution of[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (0.36 g, 0.9 mmol) in dry DMF (9 mL) was addedcesium carbonate (1.04 g, 3.19 mmol) followed by neat methyl iodide(0.064 mL, 1.03 mmol). The mixture was stirred at room temperature for3.5 hr, extracted into ethyl acetate (150 mL), washed with water (2×50mL) and brine (50 mL), dried over sodium sulfate, filtered, concentratedin vacuo and dried to give 0.34 g (93%) of pure[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester. RP-HPLC: 5.89 min; ES-MS (M+H)⁺=414.0; 1H-NMR(DMSO-d₆) δ (ppm): 1.33 (s, 9), 3.20 (s, 3), 6.75 (d, 1), 7.52 (d, 1),7.80 (d, 2), 7.83 (d, 1), 7.93 (d, 1), 8.35 (d, 2).

Example 9[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicAcid Tert-Butyl Ester

To a suspension of[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (0.33 g, 0.79 mmol) in ethyl acetate (6 mL) andethanol (2 mL) under Ar was added 10% Pd/C (0.13 g, 0.12 mmol Pd). Themixture was hydrogenated under 1 atm H₂ for 2 hr, filtered throughCelite and concentrated to give 0.28 g (92%) of[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester. RP-HPLC: 3.83 min; ES-MS (M+H)⁺=384.0; 1H-NMR(DMSO-d₆) δ (ppm): 1.32 (s, 9), 3.17 (s, 3), 5.31 (br s, 2), 6.60 (m,3), 7.00 (d, 2), 7.32 (d, 2), 7.75 (d, 1), 7.86 (d, 1).

Example 105-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide

To a suspension of 5-chlorothiophene-2-sulfonamide (0.17 g, 0.84 mmol)and N,N′-disuccinimidyl carbonate (DSC, 0.23 g, 0.91 mmol) indichloromethane (5 mL) was added tetramethylguanidine (TMG, 0.19 mL).The resulting solution was stirred at room temperature for 15 hr. Thereaction was concentrated and a solution of[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (0.27 g, 0.7 mmol) in acetonitrile (5 mL) wasadded. The resulting solution was stirred at 70° C. for 9 hr. Thereaction was diluted with dichloromethane, washed with 0.5 N HCl, driedover sodium sulfate and concentrated to give 0.48 g of crudesulfonylurea.

To a chilled solution of the crude product in dichloromethane (6 mL) andtriethylsilane (2 mL) was added neat trifluoroacetic acid (6 mL). Afterstirring at room temp for 1 hr, the reaction was concentrated,azeotroped with heptane and dried under high vac to give 0.65 g of crude5-chloro-N-[[[4-(7-chloro-6-methylamino-1-oxo-2(1H)-isoquinolinyl)phenyl]amino]carbonyl]-2-thiophenesulfonamide.This crude material was triturated with acetonitrile (5 mL), chilled andfiltered to give 0.22 g (63%) of pure5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamideas a white solid. RP-HPLC: 5.18 min; ES-MS (M+H)⁺=507.0; 1H-NMR(DMSO-d₆) δ (ppm): 2.79 (s, 3), 6.50 (d, 1), 6.53 (br s, 1), 6.70 (d,1), 7.24 (m, 2), 7.30 (d, 2), 7.46 (d, 2), 7.64 (m, 2).

Example 112-(4-Amino-phenyl)-6-cyclopropylamino-7-fluoro-2H-isoquinolin-1-one

An analogous C—N coupling procedure to that described in Example 7 wasperformed on trifluoro-methanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(Example 6) using cyclopropylamine as the nucleophile. Reduction of thenitro group was effected using the procedure outlined in Example 9.ES-MS (M+H)⁺=310.

Example 12 (5-Chloro-thiophene-2-sulfonyl)-carbamic Acid Ethyl Ester

To a solution of 5-Chloro-thiophene-2-sulfonic acid amide (4.0 g, 20.2mmol) in dry THF (200 mL) was added cesium carbonate (9.9 g, 30.3 mmol)and ethyl chloroformate (2.9 mL, 30.3 mmol). The mixture was stirred atroom temperature for 48 h. The product was taken up in H₂O (150 mL) andwashed with EtOAc (100 mL). The aqueous layer was acidified to pH=3 with1N HCl (90 mL) and the product extracted with EtOAc (100 mL). Theorganic layer was washed with brine (100 mL), dried over Na₂SO₄ andconcentrated to give a dense clear oil which solidified upon standing togive 4.41 g (81%) of 5-Chloro-thiophene-2-sulfonyl)-carbamic acid ethylester. RP-HPLC: 4.45 min. 1H-NMR (CDCl₃) δ (ppm): 7.63 (d, J=4, 1H),7.44 (bs, 1H), 6.95 (d, J=4, 1H), 4.20 (q, J=7, 2H), 1.27 (t, J=7, 3H).

Example 13

A mixture of2-(4-Amino-phenyl)-6-cyclopropylamino-7-fluoro-2H-isoquinolin-1-one(Example 11) (23 mg, 0.073 mmol) and(5-Chloro-thiophene-2-sulfonyl)-carbamic acid ethyl ester (Example 12)(28 mg, 0.10 mmol, 1.35 eq) in dry toluene (1.5 mL) was heated at 110°C. for 2 hr. Upon cooling, the reaction was concentrated in vacuo andthe crude residue was purified by HPLC (C-18) to give 17 mg (46%) ofpure5-chloro-N-[({4-[6-(cyclopropylamino)-7-fluoro-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=533, 535 (Cl).

Example 14

An analogous C—N coupling procedure to that described in Example 7 wasperformed on trifluoro-methanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(Example 6) using formamide as the nucleophile. Reduction of the nitrogroup was effected using the procedure outlined in Example 9. Couplingto form the sulfonyl urea was achieved using the method described inExample 13 to give5-chloro-N-[({4-[7-fluoro-6-(formylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=521, 523 (Cl).

Example 15

An analogous C—N coupling procedure to that described in Example 7 wasperformed on trifluoro-methanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(Example 6) using acetamide as the nucleophile. Reduction of the nitrogroup was effected using the procedure outlined in Example 9. Couplingto form the sulfonyl urea was achieved using the method described inExample 13 to giveN-(2-{4-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]phenyl}-7-fluoro-1-oxo-1,2-dihydroisoquinolin-6-yl)acetamide.ES-MS (M+H)⁺=535, 537 (Cl).

Example 16

An analogous C—N coupling procedure to that described in Example 7 wasperformed on Trifluoro-methanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(Example 6) using N-methylformamide as the nucleophile, followed byalkylation and reduction of the nitro group using the procedure outlinedin Examples 8 and 9. Coupling to form the sulfonyl urea was achievedusing the method described in Example 13 to give5-chloro-N-[({4-[7-fluoro-6-[formyl(methyl)amino]-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=535, 537 (Cl).

Example 17

An analogous C—N coupling procedure to that described in Example 7 wasperformed on trifluoro-methanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(Example 6) using N-methylacetamide as the nucleophile, followed byalkylation and reduction of the nitro group using the procedure outlinedin Examples 8 and 9. Coupling to form the sulfonyl urea was achievedusing the method described in Example 13 to giveN-(2-{4-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]phenyl}-7-fluoro-1-oxo-1,2-dihydroisoquinolin-6-yl)-N-methylacetamide.ES-MS (M+H)⁺=549, 551 (Cl).

Example 18

An analogous C—N coupling procedure to that described in Example 7 wasperformed on trifluoro-methanesulfonic acid7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl ester(Example 6) using morpholine as the nucleophile. Reduction of the nitrogroup was effected using the procedure outlined in Example 9. Couplingto form the sulfonyl urea was achieved using the method described inExample 13 to give5-chloro-N-({[4-(7-fluoro-6-morpholin-4-yl-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=563, 565 (Cl).

Example 149

An analogous alkylation procedure to that described in Example 8 wasperformed on[7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (Example 7) using ethyl iodide as the alkylatingagent. Reduction of the nitro group and coupling to form the sulfonylurea was achieved using the method described in Exs. 9 and 10, resp. togive5-chloro-N-[({4-[6-(ethylamino)-7-fluoro-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=521, 523 (Cl).

Example 20

An analogous alkylation procedure to that described in Example 8 wasperformed on[7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (Example 7) using 1-bromo-2-fluoroethane as thealkylating agent. Reduction of the nitro group and coupling to form thesulfonyl urea was achieved using the method described in Exs. 9 and 10,resp. to give5-chloro-N-[({4-[7-fluoro-6-[(2-fluoroethyl)amino]-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=539, 541 (Cl).

Example 21

An analogous alkylation procedure to that described in Example 8 wasperformed on[7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (Example 7) using2,2,2-trifluoroethyl-p-toluenesulfonate as the alkylating agent withadditional heating at 95° C. Reduction of the nitro group and couplingto form the sulfonyl urea was achieved using the method described inExs. 9 and 10, respectively, to give5-chloro-N-[({4-[7-fluoro-1-oxo-6-[(2,2,2-trifluoroethyl)amino]isoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=575, 577 (Cl).

Example 22

An analogous alkylation procedure to that described in Example 8 wasperformed on[7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (Example 7) using proparyl bromide as thealkylating agent. Reduction of the nitro group and coupling to form thesulfonyl urea was achieved using the method described in Exs. 9 and 10,resp. to give5-chloro-N-[({4-[7-fluoro-1-oxo-6-(prop-2-ynylamino)isoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=531, 533 (Cl).

Example 23

An analogous alkylation procedure to that described in Example 8 wasperformed on[7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (Example 7) using 4-chlorobenzylbromide as thealkylating agent. Reduction of the nitro group and coupling to form thesulfonyl urea was achieved using the method described in Exs. 9 and 10,resp. to give5-chloro-N-[({4-[6-[(4-chlorobenzyl)amino]-7-fluoro-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=617, 619 (2Cl).

Example 24

The alkylated intermediate synthesized in Example 23 was treated withTFA and methylated with methyl iodide and cesium carbonate. Reduction ofthe nitro group and coupling to form the sulfonyl urea was achievedusing the method described in Example 9 and 10, to give5-chloro-N-[({4-[6-[(4-chlorobenzyl)(methyl)amino]-7-fluoro-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=631, 633 (2Cl).

Example 25

Reduction of[7-fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (Example 7) was effected using the procedureoutlined in Example 9. Coupling to form the sulfonyl urea was achievedusing the method described in Example 10 to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=493, 495 (Cl).

Example 26

To a suspension of the sulfonyl urea from Example 25 (11 mg, 0.023 mmol)in glacial acetic acid (0.9 mL) was added formaldehyde (37 wt % inwater) (12 uL, 0.16 mmol) followed by sodium triacetoxyborohydride (11mg, 0.052 mmol). The reaction mixture was stirred at room temperatureovernight and concentrated in vacuo. The crude residue was purified byHPLC to give5-chloro-N-[({4-[6-(dimethylamino)-7-fluoro-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=521, 523 (Cl).

Example 27

An analogous coupling procedure described in Example 10 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) using commercially availablethiophene-2-sulfonamide to giveN-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide. ES-MS (M+H)⁺=473.

Example 28

An analogous coupling procedure as that described in Example 10 wasperformed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and commercially available5-bromothiophene-2-sulfonamide to give5-bromo-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=551, 553 (Br).

Example 29

To a solution of triphosgene (9 mg, 31 μmol) in dichloromethane (0.2 mL)was slowly added a solution of[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (30 mg, 78 μmol) and DIEA (27 μL, 156 μmol) indichloromethane (1.0 mL). The mixture was stirred at room temperaturefor 15 min. To this solution was then quickly added a solution of5-methylthiophene-2-sulfonamide (28 mg, 156 μmol) and DIEA (27 μL, 156μmol) in dichloromethane (1.0 mL). The mixture was then stirred at roomtemperature for 15 min. The reaction mixture was then diluted withdichloromethane, washed with 0.5 N HCl, dried over sodium sulfate andconcentrated to give 62 mg of crude sulfonylurea as a cloudy oil. Thecrude mixture was dissolved in TFA, reacted at room temperature for 15min., and concentrated in vacuo to give the crude final product asyellow oil which was purified by HPLC to give 17 mg (45%) ofN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-methylthiophene-2-sulfonamideas a white solid. ES-MS (M+H)⁺=487.1; 1H-NMR (DMSO-d₆) δ (ppm):8.84-8.80 (bd, J=4.5 Hz, 1H), 7.66-7.60 (d, J=12.8 Hz, 1H), 7.50-7.44(d, J=8.8 Hz, 2H), 7.40-7.36 (bs, 1H), 7.25-7.15 (m, 3H), 6.80-6-74 (bs,1H), 6.74-6.66 (d, J=8.4 Hz, 1H), 6.56-6.48 (bs, 1H), 6.50-6.46 (d, J=8Hz, 1H), 2.82-2.77 (bd, J=4.5 Hz, 3H).

Example 30

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 5-ethylthiophene-2-sulfonamide togiveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-ethylthiophene-2-sulfonamide.ES-MS (M+H)⁺=501.1.

Example 31

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 5-propylthiophene-2-sulfonic acidamide to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-propylthiophene-2-sulfonamide.ES-MS (M+H)⁺=515.1; 1H-NMR (DMSO-d₆) δ (ppm): 9.06-9.00 (s, 1H),7.66-7.62 (d, J=12.4 Hz, 1H), 7.62-7.58 (d, J=3.7 Hz, 1H), 7.48-7.42 (m,2H), 7.32-7.26 (m, 2H), 7.24-7.20 (d, J=7.3 Hz, 1H), 6.96-6-90 (d, J=3.6Hz, 1H), 6.74-6.66 (d, J=8.4 Hz, 1H), 6.58-6.48 (bs, 1H), 6.50-6.46 (d,J=7.3 Hz, 1H), 2.84-2.76 (m, 5H), 1.68-1.56 (tq, J=7.3, 7.6 Hz, 2H),0.94-0.86 (t, J=7.3 Hz, 3H).

Example 32

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and5-difluoromethyl-thiophene-2-sulfonic acid amide to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-difluoromethylthiophene-2-sulfonamide.ES-MS (M+H)⁺=523.1; 1H-NMR (DMSO-d₆) δ (ppm): 8.70-8.64 (s, 1H),7.66-7.60 (d, J=12.4 Hz), 7.56-7.48 (m, 2h), 7.38-7.10 (t, J=55.3 Hz,1H), 7.36-7.32 (m, 1H), 7.28-7.24 (m, 1H), 7.22-7.18 (d, J=7.7 Hz, 1H),7.12-7.06 (m, 2H), 6.72-6.66 (d, J=8.4 Hz, 1H), 6.52-6.46 (bs, 1H),6.48-6.44 (d, J=7.3 Hz, 1H), 2.81-2.77 (bd, J=4.7 Hz, 3H).

Example 33

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 5-cyanothiophene-2-sulfonamide togiveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-cyanothiophene-2-sulfonamide.ES-MS (M+H)⁺=498.1.

Example 34

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 5-methoxythiophene-2-sulfonamideto giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-ethylthiophene-2-sulfonamide.ES-MS (M+H)⁺=503.1; 1H-NMR (DMSO-d₆) δ (ppm): 9.15-9.00 (s, 1H),7.68-7.60 (d, J=12.5 Hz, 1H), 7.52-7.50 (d, J=3.3 Hz, 1H), 7.48-7.44 (d,J=8.8 Hz, 2H), 7.34-7.28 (d, J=8.8 Hz, 2H), 7.25-7.20 (d, J=7.3 Hz, 1H),6.74-6.68 (d, J=7.7 Hz, 1H), 6.56-6.50 (bs, 1H), 6.52 (d, J=7.7 Hz, 1H),6.44-6.40 (d, J=3.3 Hz, 1H), 3.94-3.92 (s, 3H), 2.82-2.78 (bd, J=4.4 Hz,3H).

Example 35

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 5-ethynylthiophene-2-sulfonamideto giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-ethynylthiophene-2-sulfonamide.ES-MS (M+H)⁺=497.1; 1H-NMR (DMSO-d₆) δ (ppm): 9.22-9.18 (bs, 1H),7.70-7.66 (m, 1H), 7.65-7.60 (d, J=12.4 Hz, 1H), 7.48-7.42 (d, J=9.1 Hz,2H), 7.40-7.36 (m, 1H), 7.32-7.25 (d, J=8.8 Hz, 2H), 7.24-7.18 (d, J=7.7Hz, 1H), 6.72-6.66 (d, J=8.4 Hz, 1H), 6.58-6.48 (bs, 1H), 6.50-6.46 (d,J=7.3 Hz, 1H), 4.86-4.84 (s, 1H), 2.82-2.76 (s, 3H).

Example 36

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and5-carboxamidethiophene-2-sulfonamide to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-5-carboxamidethiophene-2-sulfonamide.ES-MS (M+H)⁺=516.1.

Example 37

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 4-chlorothiophene-2-sulfonamide togiveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-4-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=507.0, 509.0 (Cl).

Example 38

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 4-methylthiophene-2-sulfonamide togiveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-4-methylthiophene-2-sulfonamide.ES-MS (M+H)⁺=487.1.

Example 39

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and4-difluoromethylthiophene-2-sulfonamide to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-4-difluoromethylthiophene-2-sulfonamide.ES-MS (M+H)⁺=523.1; 1H-NMR (DMSO-d₆) δ (ppm): 9.24-9.16 (bs, 1H),8.34-8.28 (m, 1H), 7.88-7.84 (m, 1H), 7.66-7.60 (d, J=12.4 Hz, 1H),7.48-7.42 (d, J=8.8 Hz, 2H), 7.34-7.26 (d, J=8.8 Hz, 2H), 7.25-7.20 (d,J=7.3 Hz, 1H), 7.19-6.91 (t, J=55.3 Hz, 1H), 6.72-6.66 (d, 8.4 Hz, 1H),6.58-6.48 (bs, 1H), 6.50-6.46 (d, J=7.3 Hz, 1H), 2.82-2.77 (s, 3H).

Example 40

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) using 3-chloro-thiophene-2-sulfonicacid amide as the coupling partner to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-3-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=507.0, 509.0 (Cl); 1H-NMR (DMSO-d₆) δ (ppm): 8.98-8.91 (bs,1H), 8.06-7.98 (m, 1H), 7.66-7.60 (d, J=12.8 Hz, 1H), 7.48-7.40 (d,J=8.8 Hz, 2H), 7.30-7.25 (d, J=8.8 Hz, 2H), 7.24-7.20 (m, 1H), 7.24-7.18(d, J=7.3 Hz, 1H), 6.72-6.66 (d, J=8.4 Hz, 1H), 6.56-6.48 (bs, 1H),6.50-6.46 (d, J=7.3 Hz, 1H), 2.82-2.75 (s, 3H).

Example 41

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and 3-methyl-thiophene-2-sulfonamideto giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-3-methylthiophene-2-sulfonamide.ES-MS (M+H)⁺=487.1; 1H-NMR (DMSO-d₆) δ (ppm): 8.96-8.92 (bs, 1H),7.86-7.82 (d, J=5.1 Hz, 1H), 7.66-7.60 (d, J=12.4 Hz, 1H), 7.46-7.40 (m,2H), 7.32-7.26 (m, 2H), 7.24-7.20 (d, J=7.3 Hz, 1H), 7.03-7.00 (d, J=5.1Hz, 1H), 6.72-6.68 (d, J=8.8 Hz, 1H), 6.56-6.48 (bs, 1H), 6.50-6.46 (d,J=7.7 Hz, 1H), 2.82-2.78 (bd, J=4.8 Hz, 3H), 2.45-2.43 (s, 3H).

Example 42

An analogous sulfonylurea coupling and de-protection procedure to thatdescribed in Example 29 was performed on[2-(4-Amino-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (Example 9) and benzenesulfonamide to giveN-({[4-(6-amino-7-fluoro-1-oxoisoquinolin-2(1H)-yl)phenyl]amino}carbonyl)-benzenesulfonamide.ES-MS (M+H)⁺=467.1; 1H-NMP (ΔMΣO-66) δ (ppm): 9.13-9.06 (bs, 1H),7.98-7.92 (d, J=8.4 Hz, 2H), 7.70-7.56 (m, 4H), 7.44-7.37 (d, J=8.8 Hz,2H), 7.39-7.24 (d, J=8.8 Hz, 2H), 7.21-7.18 (d, J=7.3 Hz, 1H), 6.74-6.66(d, J=8.4 Hz, 1H), 6.56-6.48 (bs, 1H), 6.49-6.46 (d, J=7.7 Hz, 1H),2.82-2.76 (bd, J=4.4 Hz, 3H).

Example 43 Trifluoromethanesulfonic Acid7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl Ester

To a chilled suspension of 7-Fluoro-6-methoxy-2H-isoquinolin-1-one (fromExample 3) (9.65 g, 50 mmol) in dichloromethane (200 mL) was added neatboron tribromide (21 mL, 220 mmol) via syringe. The yellow suspensionwas stirred at room temperature for 17 hr. The reaction was slowlypoured into methanol (300 mL) on an ice bath. The resulting solution wasconcentrated in vacuo, washed and concentrated several times withmethanol and dichloromethane, and dried to give 13 g of crude phenol.1H-NMR (DMSO-d₆) δ (ppm): 6.38 (d, 1), 7.04 (m, 2), 7.73 (d, 1), 11.05(s, 1).

To a suspension of 10.7 g of crude phenol in pyridine (160 mL) was addedDMAP (7.6 g, 62.3 mmol) followed by phenyltrifluoromethylsulfonimide(17.4 g, 48.6 mmol) portionwise over approx. 5 min. The reaction mixturewas stirred at room temperature for 1.5 hr, extracted into ethyl acetate(600 mL), washed with water (3×250 mL) and brine (250 mL). The organiclayer was dried over sodium sulfate, filtered, concentrated and dried togive 24 g crude product, which was triturated withdichloromethane/hexane (2:1) to give 11.6 g (90% yield for 2 steps) oftrifluoromethane-sulfonic acid7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl ester. 1H-NMR (DMSO-d₆) δ(ppm): 6.63-6.65 (s, 1), 7.26-7.29 (t, 1), 8.11-8.16 (m, 2), 11.61 (brs, 1).

Example 44[7-Fluoro-1-oxo-2-(2-trimethylsilanyl-ethoxymethyl)-1,2-dihydro-isoquinolin-6-yl]-carbamicAcid Tert-Butyl Ester

To a solution of trifluoromethane-sulfonic acid7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl ester (10.5 g, 33.8 mmol) inTHF (155 mL) at 0° C. was added neat 2-(trimethylsilyl)-ethoxymethylchloride (SEM-Cl, 15 mL, 85 mmol) dropwise over 5 min, followed by neatDBU (19 mL, 127 mmol). The reaction slurry was stirred at roomtemperature for 2 hr, diluted with ethyl acetate (600 mL), washed with0.25N HCl (200 mL), water (200 mL) and brine (250 mL), dried overNa₂SO₄, filtered, concentrated and dried to give crude product. Silicagel chromatography using 5-20% ethyl acetate/hexane as eluent gave 9.2 g(62%) of pure SEM-protected triflate. 1H-NMR (DMSO-d₆) δ (ppm): 0.085(s, 9), 0.81-0.85 (d, 2), 3.53-3.57 (t, 2), 5.32 (s, 2), 6.72-6.74 (d,1), 7.56-7.58 (d, 1), 8.12-8.14 (d, 1), 8.20-8.23 (d, 1).

In a dry flask was combined the triflate (9.2 g, 21 mmol), t-butylcarbamate (3.42 g, 29.2 mmol), dry powdered cesium carbonate (11.3 g,34.7 mmol), 9,9-dimethyl-4,5-bis(diphenylphosphino)xanthene (Xantphos,1.45 g, 2.5 mmol), and tris(dibenzylideneacetone)dipalladium(0) (Pd₂dba₃, 0.38 g, 0.83 mmol Pd). Under Ar atmosphere, dry THF (140 mL) wasadded to the flask, and the mixture was stirred at 70° C. for 3 hr. Uponcooling, the reaction was diluted with hexane (80 mL), filtered andconcentrated to give 9.0 g crude product, which was purified by columnchromatography (silica, 10-25% EtOAc/hexane) to give 6.1 g (72%) of pure[7-Fluoro-1-oxo-2-(2-trimethylsilanyl-ethoxymethyl)-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester. ES-MS (M+H)⁺=409; 1H-NMR (DMSO-d₆) δ (ppm): −0.10(s, 9), 0.80-0.84 (t, 2), 1.46 (s, 9), 3.51-3.55 (t, 2), 5.27 (s, 2),6.58-6.60 (d, 1), 7.36-7.38 (d, 1), 7.82-7.85 (d, 1), 8.05-8.07 (d, 1),9.45 (s, NH).

Example 45 (7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamicAcid Tert-Butyl Ester

To a solution of[7-Fluoro-1-oxo-2-(2-trimethyl-silanyl-ethoxymethyl)-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (5.95 g, 14.6 mmol) in dry dimethylformamide (50mL) was added powdered cesium carbonate (12 g, 36.8 mmol) followed byneat methyl iodide (0.95 mL, 15.2 mmol). After 1 hr at room temperaturethe reaction was diluted with ethyl acetate (400 mL), washed with water(2×100 mL) and brine (100 mL), dried over Na₂SO₄, filtered, concentratedand dried to give crude methylated product. This crude material wasdissolved in 1M tetrabutylammonium fluoride (85 mL in THF) and stirredat 65° C. for 2 hr. The reaction mixture was extracted into ethylacetate (400 mL), washed with dilute HCl (100 mL), water (100 mL) andbrine (100 mL), and dried to give a crude product, which was purified bycolumn chromatography (silica, 40-70% EtOAc/dichloromethane) to give3.34 g (78%) of pure(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester. ES-MS (M+H)⁺=293.1; 1H-NMR (DMSO-d₆) δ (ppm): 1.32 (s,9), 3.17 (s, 3), 6.50-6.52 (d, 1), 7.13-7.16 (t, 1), 7.70-7.72 (d, 1),7.81-7.84 (d, 1), 11.35 (br s, 1).

Example 46

Method A: Using Substituted 4-fluoronitrobenzenes.

To a solution of(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester (Example 45) (70 mg, 0.24 mmol) and3-chloro-4-fluoronitrobenzene (55 mg, 0.31 mmol, 1.3 eq) in drydimethylformamide (2 mL) was added powdered cesium carbonate (0.2 g, 0.6mmol, 2.5 eq). The mixture was stirred vigorously at 65-70° C. for 5 hr,then chilled on an ice bath. Addition of water precipitated out thedesired product which upon filtration and drying under high vacuum gave95 mg (89%) of pure nitro-aryl product. 1H-NMR (DMSO-d₆) δ (ppm): 1.34(s, 9), 3.21 (s, 3), 6.76-6.78 (d, 1), 7.37-7.39 (d, 1), 7.84-7.85 (d,1), 7.91-7.93 (d, 2), 8.34-8.36 (dd, 1), 8.54-8.55 (d, 1).

This nitro intermediate (89 mg, 0.2 mmol) was reduced by reaction withtin (II) dichloride dihydrate (134 mg, 0.6 mmol, 3 eq) in ethanol (3 mL)at 70° C. for 2 hr. Upon cooling, the reaction was diluted with ethylacetate (20 mL), treated with Celite and 5% sodium bicarbonate (10 mL)to precipitate the tin (II) oxide. The Celite/SnO₂ was filtered off, andthe organic layer washed with 5% NaHCO₃ (10 mL) and brine (10 mL), driedover Na₂SO₄, conc. in vacuo to gave 83 mg (100%) of the desired aniline[2-(4-Amino-2-chloro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester. ES-MS (M+H)⁺=418, 420 (Cl).

Example 47

Method B: Using Substituted 4-halo-nitrobenzenes or2-halo-5-nitropyridines.

In a dry flask was combined(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester (Example 45) (58 mg, 0.2 mmol), 2-bromo-5-nitropyridine(61 mg, 0.3 mmol, 1.5 eq), dry powdered cesium carbonate (113 mg, 0.35mmol, 1.73 eq), 9,9-dimethyl-4,5-bis(diphenylphosphino)-xanthene(Xantphos, 15 mg, 0.026 mmol), andtris(dibenzylideneacetone)dipalladium(0) (Pd₂ dba₃, 4.5 mg, 0.01 mmolPd). Under Ar atmosphere, dry THF (2 mL) was added to the flask, and themixture was stirred at 80° C. for 2 hr. Upon cooling, the reaction wasconcentrated and the crude residue was purified by column chromatography(silica 2-15% EtOAc/dichloromethane) to give 68 mg (83%) of purenitro-pyridyl product. 1H-NMR (DMSO-d₆) δ (ppm): 1.35 (s, 9), 3.21 (s,3), 6.81-6.83 (d, 1), 7.84-7.85 (d, 1), 7.88-7.90 (d, 1), 7.99-8.01 (d,1), 8.24-8.26 (d, 1), 8.76-8.79 (dd, 1), 9.39-9.40 (d, 1).

This nitro intermediate (67 mg, 0.16 mmol) was reduced under catalytichydrogenation conditions using 1 atm H₂, 10% Pd/C (26 mg, 0.024 mmol Pd)in ethanol (2 mL) for 3 hr to give 60 mg (97%) of[2-(5-Amino-pyridin-2-yl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester. ES-MS (M+H)⁺=385.

Example 48

Method C: Using Substituted 4-haloanilines or 5-halo-2-aminopyridinesand Pyrimidines

In a dry flask was combined(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester (Example 45, compound 10, Scheme B) (102 mg, 0.35mmol), 2-amino-5-iodopyridine (84 mg, 0.38 mmol, 1.1 eq), copper (I)iodide (7 mg, 0.037 mmol, 0.11 eq), 8-hydroxyquinoline (6 mg, 0.041mmol, 0.12 eq) and powdered potassium carbonate (58 mg, 0.42 mmol, 1.2eq). Under Ar atmosphere, dry dimethylsulfoxide (DMSO, 1.5 mL) wasadded, and the mixture was stirred at 115° C. for 50 hr. The reactionwas cooled, concentrated and purified by chromatography (silica, 2-15%isopropyl alcohol/dichloromethane) to give 67 mg (50%) of[2-(6-Amino-pyridin-3-yl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester as a grey solid. ES-MS (M+H)⁺=385. 1H-NMR (DMSO-d)δ (ppm): 1.33 (s, 9), 3.19 (s, 3), 6.25 (br m, NH₂), 6.53 (m, 1),6.64-6.65 (d, 1), 7.37-7.39 (d, 1), 7.44-7.46 (d, 1), 7.77-7.79 (d, 1),7.87-7.90 (d, 1), 7.95 (m, 1H).

Example 49

An analogous procedure to that outlined in Example 46 (Method A) using3,4-difluoronitrobenzene was used to prepare the intermediate aniline.Formation of the sulfonyl urea was achieved using the method describedin Example 13, followed by TFA de-protection, to give5-chloro-N-[({3-fluoro-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=525, 527 (Cl).

Example 50

An analogous procedure to that outlined in Example 46 (Method A) using3-chloro-4-fluoronitrobenzene, was used to prepare the intermediateaniline. Formation of the sulfonyl urea was achieved using the methoddescribed in Example 13, followed by TFA de-protection, to give5-chloro-N-[({3-chloro-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=541, 543 (2Cl).

Example 51

An analogous procedure to that outlined in Example 46 (Method A) using3-bromo-4-fluoronitrobenzene was used to prepared the intermediateaniline. Formation of the sulfonyl urea was achieved using the methoddescribed in Example 13, followed by TFA de-protection, to giveN-[({3-bromo-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=585, 587, 589 (BrCl).

Example 52

An analogous procedure to that outlined in Example 46 (Method A) using4-fluoro-3-methylnitrobenzene was used to afford the substitutedaniline. Formation of the sulfonyl urea was achieved using the methoddescribed in Example 13, followed by TFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-methylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=520, 522 (Cl).

Example 53

An analogous procedure to that outlined in Example 46 (Method A) using4-fluoro-2-methylnitrobenzene was used to prepare the substitutedaniline. Formation of the sulfonyl urea was achieved using the methoddescribed in Example 13, followed by TFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-methylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=520, 522 (Cl).

Example 54

An analogous procedure to that outlined in Example 47 (Method B) using2-bromo-5-nitropyridine was used to prepare the substitutedaminopyridine. Formation of the sulfonyl urea was achieved using themethod described in Example 13, followed by TFA deprotection, to give5-chloro-N-[({6-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]pyridin-3-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=508, 510 (Cl).

Example 55

An analogous procedure to that outlined in Example 47 (Method B) using2-chloro-5-nitro-6-methylpyridine was used to prepare the substitutedaminopyridine. Formation of the sulfonyl urea was achieved using themethod described in Example 13, followed by TFA deprotection, to give5-chloro-N-[({6-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-methylpyridin-3-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=522, 524 (Cl).

Example 56

An analogous procedure to that outlined in Example 47 (Method B) using2-chloro-4-methyl-5-nitropyridine was used to prepare the substitutedaminopyridine. Formation of the sulfonyl urea was achieved using themethod described in Example 13, followed by TFA deprotection, to give5-chloro-N-[({6-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-4-methylpyridin-3-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=522, 524 (Cl).

Example 57

An analogous procedure to that outlined in Example 47 (Method B) using2-chloro-3-methyl-5-nitropyridine was used to prepare the substitutedaminopyridine. Formation of the sulfonyl urea was achieved using themethod described in Example 13, followed by TFA deprotection, to give5-chloro-N-[({6-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-5-methylpyridin-3-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=522, 524 (Cl).

Example 58

An analogous procedure to that outlined in Example 48 (Method C) using2-fluoro-4-iodoaniline was used to prepare the substituted aniline.Formation of the sulfonyl urea was achieved using the method describedin Example 13, followed by TFA deprotection, to give5-chloro-N-[({2-fluoro-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=525, 527 (Cl).

Example 59

An analogous procedure to that outlined in Example 48 (Method C) using2-amino-5-bromopyridine was used to prepare the intermediateaminopyridine. Formation of the sulfonyl urea was achieved using themethod described in Example 13, followed by TFA deprotection, to give5-chloro-N-[({5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]pyridin-2-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=508, 510 (Cl).

Example 60

An analogous procedure to that outlined in Example 48 (Method C) using2-amino-3-methyl-5-bromopyridine was used to prepare the substitutedaniline. Formation of the sulfonyl urea was achieved using the methoddescribed in Example 13, followed by TFA deprotection, to give5-chloro-N-[({5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-methylpyridin-2-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=521, 523 (Cl).

Example 61

An analogous procedure to that outlined in Example 48 (Method C) using2-amino-5-iodopyrimidine was used to prepare the intermediateaminopyrimidine. Formation of the sulfonyl urea was achieved using themethod described in Example 13, followed by TFA deprotection, to give5-chloro-N-[({5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]pyrimidin-2-yl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=509, 511(Cl).

Example 62

To a solution of 2-fluoro-5-nitrobenzyl alcohol (1 g, 5.84 mmol) in 5 mLdichloromethane and triethylamine (0.81 ml, 5.84 mmol), was added acetylchloride (0.415 ml, 1 eq) dropwise. The solution was stirred at roomtemperature for 12 hours, diluted with ethyl acetate and extracted withbrine. Combine organic layers was dried over sodium sulfate,concentrated in vacuo to give acetic acid 2-fluoro-5-nitro-benzyl ester.

Acetic acid 2-fluoro-5-nitro-benzyl ester was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-2-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzylacetate. ES-MS (M+H)⁺=579, 581(Cl).

Example 63

This analog compound was obtained from hydrolysis of the intermediate,Boc protected compound of Example 62, then followed by TFA deprotectionto give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-(hydroxymethyl)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=537, 539(Cl).

Example 64

To a solution of 2-fluoro-5-nitrobenzyl alcohol (1 g, 5.84 mmol) in 25mL ether, was added tetrabromomethane (3.87 g, 11.7 mmol), followed bytriphenylphosphine (3.39 g, 11.7 mmol). The mixture was stirred at roomtemperature for 2 hours. The reaction was concentrated and the cruderesidue purified by column chromatography (silica, 10% EtOAc/hexane) togive pure 2-bromomethyl-1-fluoro-4-nitro-benzene.

To a solution of 2-bromomethyl-1-fluoro-4-nitro-benzene (0.2 g, 0.85mmol) in 5 mL dry THF, was added piperidine (0.11 ml, 1 mmol) and DIEA(0.3 ml, 1.7 mmol) at 0° C. The resulting reaction was stirred at 0° C.to room temperature for 1 hour, then diluted with EtOAc and washed withbrine. Combined organic layer was dried over sodium sulfate,concentrated in vacuo and the crude residue was purified by columnchromatography (10% EtOAc/hexane) to give pure1-(2-fluoro-5-nitro-benzyl)piperidinebenzene.

1-(2-Fluoro-5-nitro-benzyl)piperidinebenzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-(piperidin-1-ylmethyl)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=604, 606(Cl).

Example 65

To a solution of 2-fluoro-5-nitro-benzoic acid (0.269 g, 1.45 mmol) in 3mL ethanol, was added concentrated sulfuric acid (0.5 ml). The solutionwas refluxed under argon for 3 hours. The mixture was stirred at roomtemperature for 2 hours, then diluted with ethyl acetate and washed withbrine. The organic layer was dried over sodium sulfate, concentrated invacuo to give pure 2-fluoro-5-nitro-benzoic acid ethyl ester.

2-Fluoro-5-nitro-benzoic acid ethyl ester was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give ethyl5-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-2-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzoate.ES-MS (M+H)⁺=579, 581(Cl).

Example 66

This analog compound was obtained from hydrolysis of ethyl5-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-2-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzoateof Example 65 using the procedure described in Example 63.5-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-2-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzoicacid. ES-MS (M+H)⁺=551, 553(Cl).

Example 67

The substituted aniline was generated by Method A (Example 46) using2-fluoro-5-nitro-benzonitril. Formation of the sulfonyl urea wasachieved using the method described in Example 10, followed by TFAdeprotection, to give5-chloro-N-[({3-cyano-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=532, 534 (Cl).

Example 68

This analog compound was obtained during the TFA de-protection step inExample 67.5-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-2-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzamide.ES-MS (M+H)⁺=550, 552 (Cl).

Example 69

To a solution of 2-bromomethyl-1-fluoro-4-nitro-benzene (131 mg, 0.56mmol), which was generated in Example 64) and dimethylaminehydrochloride (44 mg, 0.54 mmol) in 4 mL dioxane, was added cesiumcarbonate (546 mg, 1.68 mmol). The mixture was heated to 70° C. underargon for 12 hours. Work up and RP prep HPLC to give(2-fluoro-5-nitro-benzyl)-dimethyl-amine.

(2-Fluoro-5-nitro-benzyl)-dimethyl-amine was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[(f{3-[(dimethylamino)methyl]-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=564, 566(Cl).

Example 70

To a solution of 3-bromo-4-fluoronitrobenzene (696 mg, 3.16 mmol) andtributyl(1-ethoxyvinyl)tin (1.07 ml, 3.5 mmol) in 10 mL toluene, wasadded tetrakis(triphenylphosphine)palladium(0) (183 mg, 016 mmol). Themixture was purged with argon for 3 minutes, and then heated to 110° C.under argon for 2 days. The reaction mixture was cooled to roomtemperature, 3 mL 1 N HCl was added and stirred at room temperature for40 minutes. Workup followed by column chromatographic purification(silica 10%-30% EtOAc/hexane) to give pure1-(2-fluoro-5-nitro-phenyl)-ethanone.

1-(2-fluoro-5-nitro-phenyl)-ethanone was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to giveN-[({3-acetyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide.ES-MS(M+H)⁺=549, 551(Cl).

Example 71

A mixture of bis(benzonitril)dichloropalladium(II) (53 mg, 0.14 mmol)and copper (I) iodide (26 mg, 0.14 mmol) in 6 mL dry THF was purged withargon for 3 minutes, then was added tri(tert-butyl phosphine) (69 ul,0.28 mmol), TMS acetylene (0.77 ml, 5.5 mmol),3-bromo-4-fluoronitrobenzene 505 mg, 2.295 mmol), and di-isopropylamine(0.77 ml, 5.5 mmol). The mixture was stirred at room temperature for 5hours under argon. To the reaction mixture was added 7 mL 1 M TBAF/THFand the mixture was stirred at room temperature for 10 minutes. Workupand purification by column chromatography (silica, 5%-35% EtOAc/hexane)give 2-ethynyl-1-fluoro-4-nitro-benzene.

2-Ethynyl-1-fluoro-4-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({3-ethynyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=531, 533(Cl).

Example 72

To a solution of 2-ethynyl-1-fluoro-4-nitro-benzene (70 mg, 0.42 mmol,obtained in Example 71) in 3 mL ethanol, 5% Pd/BaSO4 (48 mg) was added.The mixture was hydrogenated at 1 atm for 1.5 hrs. Catalyst was removedby filtering through a celite pad. The filtrate was concentrated to givepure 1-fluoro-4-nitro-2-vinyl-benzene.

1-Fluoro-4-nitro-2-vinyl-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-vinylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=533, 535(Cl).

Example 73

To a solution of[7-fluoro-2-(4-nitro-2-vinyl-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (60 mg, 0.137 mmol, the intermediate materialgenerated in Example 72) in 1.5 mL ethanol and 2.5 mL ethylacetate, 5%Pd/C (60 mg) was added and the mixture was hydrogenated at 1 atm for 10hrs. Catalyst was removed by filtering through a celite pad. Thefiltrate was concentrated to give desired aniline.

Formation of the sulfonyl urea was achieved using the method describedin Example 10, followed by TFA deprotection, to give5-chloro-N-[({3-ethyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=535, 537(Cl).

Example 74

This analog compound was obtained from reduction ofN-[({3-acetyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamideof Example 70. To a solution ofN-[({3-acetyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide(18 mg, 0.033 mmol) in 2 mL ethanol, was added sodium borohydride (20mg, 0.53 mmol). The mixture was stirred at room temperature for 30minutes. Workup and purification provided5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-(1-hydroxyethyl)phenyl]amino}carbonyl)thiophene-2-sulfonamideas a mixture of rotamers. ES-MS (M+H)⁺=551, 553(Cl).

Example 75

To a solution of 5-fluoro-2-nitro-phenol (369 mg, 2.35 mmol) in 5 mLTHF, sodium hydride (96 mg, 2.46 mmol) was added at room temperature,followed by methyl iodide (0.88 ml, 14 mmol). The mixture was stirred atroom temperature for 10 hours, and then cesium carbonate (744 mg, 2.35mmol) was added. The mixture was stirred at room temperature foradditional 4 hours, then diluted with ethyl acetate and washed withbrine. The organic layers were combined and concentrated in vacuo togive a crude residue, which was purified by column chromatography(silica 5-25% EtOAc/hexane) to give pure4-fluoro-2-methoxy-1-nitro-benzene.

4-Fluoro-2-methoxy-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-methoxyphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=537, 539(Cl).

Example 76

To a solution of 5-fluoro-2-nitro-phenol (234 mg, 1.49 mmol) in 5 mLTHF, sodium hydride (122 mg, 2.9 mmol) was added, followed bychloromethoxy methane (113 ul, 1.49 mmol) at 0° C. The mixture waswarmed to room temperature and stirred for 10 hours, then diluted withethyl acetate and washed with brine. The organic layers were combinedand concentrated in vacuo to give a crude residue, which was purified bycolumn chromatography (silica 5-25% EtOAc/hexane) to give4-fluoro-2-methoxymethoxy-1-nitro-benzene.

4-Fluoro-2-methoxymethoxy-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(methoxymethoxy)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=567, 569(C).

Example 77

This analog compound was obtained from TFA deprotection step in Example76.5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-hydroxyphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=523, 525(Cl).

Example 78

2-Ethynyl-4-fluoro-1-nitro-benzene was obtained from2-bromo-4-fluoro-1-nitro-benzene using the procedure described inExample 71.

4-Fluoro-1-nitro-2-vinyl-benzene was obtained using2-ethynyl-4-fluoro-1-nitro-benzene using the procedure described inExample 72.

4-Fluoro-1-nitro-2-vinyl-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-vinylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=533, 535(Cl).

Example 79

A mixture of 5-fluoro-2-nitro-phenol (464 mg, 2.95 mmol), benzyl bromide(0.37 ml, 3.10 mmol) and cesium carbonate (1.055 g, 3.24 mmol) in MeCN(10 mL) was stirred room temperature under argon for 48 hours. Thereaction mixture was diluted with ethyl acetate and washed with brine.The organic layers were combined and concentrated in vacuo to give acrude residue, which was purified by column chromatography (silica 5-25%EtOAc/hexane) to give 2-benzyloxy-4-fluoro-1-nitro-benzene.

2-Benzyloxy-4-fluoro-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46).

Formation of the sulfonyl urea was achieved using the method describedin Example 10, followed by TFA deprotection, to giveN-[({2-(benzyloxy)-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=613, 615(Cl).

Example 80

[2-(3-Bromo-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester was generated by Method A (Example 46) using2-bromo-4-fluoro-1-nitro-benzene. A mixture of[2-(3-Bromo-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (21 mg, 0.0427 mmol),tetrakis(triphenylphosphine)palladium(0) (2.5 mg, 0.002 mmol), copper(I) iodide (2 mg, 0.008 mmol), TMS acetylene (10 ul, 0.07 mmol),n-butylamine (1.5 ml) and 1 mL DMF was purged with argon for 2 minutes.The mixture was then subjected to microwave irradiation (120° C.) for 5minutes, then diluted with ethyl acetate and washed with brine. Theorganic layers were combined and concentrated in vacuo to give a cruderesidue, which was purified by column chromatography (silica 5-25%EtOAc/hexane) to give[2-(3-butylamino-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester.

Reduction of the nitro group, sulfonyl urea formation, followed by TFAdeprotection, providedN-[({2-(butylamino)-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=578, 580(Cl).

Example 81

This analog compound was obtained from[7-fluoro-2-(4-nitro-3-vinyl-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (intermediate in Example 78) using the reductionprocedure described in Example 73.

Formation of the sulfonyl urea was achieved using the method describedin Example 10, followed by TFA deprotection, to give5-chloro-N-[({2-ethyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=535, 537(Cl).

Example 82

Acetic acid 2-(5-fluoro-2-nitro-phenoxy)-ethyl ester was obtainedstarting with acetic acid 2-bromo-ethyl ester and using the proceduredescribed in Example 79.

Acetic acid 2-(5-fluoro-2-nitro-phenoxy)-ethyl ester was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give2-{2-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenoxy}ethylacetate. ES-MS (M+H)⁺=609, 611(Cl).

Example 83

This analog compound was obtained from hydrolysis (as described inExample 63) of the intermediate Boc protected compound of Example 82,followed by TFA deprotection.5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(2-hydroxyethoxy)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=567, 569(Cl).

Example 84

4-Fluoro-2-isopropoxy-1-nitro-benzene was obtained using5-fluoro-2-nitro-phenol and isopropanol under Mitsunobu reactioncondition. The general procedure is described as follows: to a solutionof 4-fluoro-2-isopropoxy-1-nitro-benzene (313 mg, 1.99 mmol),triphenylphosphine (783 mg, 2.98 mmol) and ispropanol (161 mg, 2.59mmol) in 2 mL THF, was added diethyl azodicarboxylate (0.49 ml, 2.99mmol) dropwise at 0° C. The mixture was warmed to room temperature andstirred for 30 minutes, then diluted with ethyl acetate and washed withbrine. The organic layers were combined and concentrated in vacuo togive a crude residue, which was purified by column chromatography(silica 5-25% EtOAc/hexane) to give4-fluoro-2-isopropoxy-1-nitro-benzene.

4-Fluoro-2-isopropoxy-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-isopropoxyphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=565, 567(Cl).

Example 85

To a solution of 5-fluoro-2-nitro-benzaldehyde (573 mg, 3.39 mmol) in 4mL DCM, was added (diethylamino)sulfurtrifluride (0.448 ml, 3.39 mmol)dropwise at 0° C. over 3 minutes. The reaction mixture was stirred at 0°C. for 1 hour, then diluted with dichloromethane and washed with brine.The organic layers were combined and concentrated in vacuo to give acrude residue, which was purified by column chromatography (silica15-40% EtOAc/hexane) to give 2-difluoromethyl-4-fluoro-1-nitro-benzene.

2-Difluoromethyl-4-fluoro-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to5-chloro-N-[({2-(difluoromethyl)-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=557, 559(Cl).

Example 86

2-Cyclopropylmethoxy-4-fluoro-1-nitro-benzene was obtained fromcyclopropyl methanol using the procedure described in Example 84.

2-Cyclopropylmethoxy-4-fluoro-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to5-chloro-N-[({2-(cyclopropylmethoxy)-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=577, 579(Cl).

Example 87

The substituted aniline was generated by Method C (Example 48) using4-bromo-2-trifluoromethoxy-phenylamine. Formation of the sulfonyl ureawas achieved using the method described in Example 10, followed by TFAdeprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(trifluoromethoxy)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=591, 593(Cl).

Example 88

1-(5-Fluoro-2-nitro-phenyl)-ethanone was obtained using2-bromo-4-fluoro-1-nitro-benzene with the procedure described in Example70.

1-(5-fluoro-2-nitro-phenyl)-ethanone was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to giveN-[({2-acetyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=549, 551(Cl).

Example 89

To a solution of trifluoroethanol in 4 mL dry THF, was addedtert-butoxide (378 mg, 3.37 mmol) at 0° C. The resulting mixture wasadded dropwise to the solution of 2,4-difluoro-1-nitro-benzene (536 mg,3.37 mmol) in 5 mL dry THF at 0° C. The mixture was stirred at 0° C. for30 minutes, then diluted with ethyl acetate and washed with brine. Theorganic layers were combined and concentrated in vacuo to give4-fluoro-1-nitro-2-(2,2,2-trifluoro-ethoxy)-benzene.

4-Fluoro-1-nitro-2-(2,2,2-trifluoro-ethoxy)-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(2,2,2-trifluoroethoxy)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=605, 607(Cl).

Example 90

2-Ethoxy-4-fluoro-1-nitro-benzene was obtained as a side product whenprepared Example 89 using 2,2,2-trifluoro-ethanol as described inExample 84.

2-Ethoxy-4-fluoro-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46).

Formation of the sulfonyl urea was achieved using the method describedin Example 10, followed by TFA deprotection, to give5-chloro-N-[({2-ethoxy-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=551, 553(Cl).

Example 91

4-Fluoro-2-(2-methoxy-ethoxy)-1-nitro-benzene was obtained from2-methoxy-ethanol using the procedure described in Example 89.

4-Fluoro-2-(2-methoxy-ethoxy)-1-nitro-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(2-methoxyethoxy)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=581, 583(Cl).

Example 92

To a solution of 2-isopropyl-phenylamine (262 mg, 1.94 mmol) and sodiumacetate (159 mg, 1.94 mmol) in 5 mL acetic acid, was added iodidemonochloride (409 mg, 2.58 mmol) at room temperature. The mixture wasstirred at room temperature for 20 minutes, then diluted with ethylacetate and washed with saturated sodium bicarbonate. The organic layerswere combined and concentrated in vacuo to give a crude residue, whichwas purified by column chromatography (silica 5-25% EtOAc/hexane) togive 4-iodo-2-isopropyl-phenylamine.

4-Iodo-2-isopropyl-phenylamine was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method C (Example 48). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-isopropylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=549, 551(Cl).

Example 93

[2-(5-Fluoro-2-nitro-phenoxy)-ethyl]-dimethyl-amine was obtained from2-dimethylamino-ethanol using the procedure described in Example 89.

[2-(5-Fluoro-2-nitro-phenoxy)-ethyl]-dimethyl-amine was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({2-[2-(dimethylamino)ethoxy]-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=594, 596(Cd).

Example 94

[2-(3-Cyclopropyl-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester was obtained from[7-fluoro-2-(4-nitro-3-vinyl-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl (intermediate in Example 78).

To a solution of[7-fluoro-2-(4-nitro-3-vinyl-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl (12 mg, 0.027 mmol) in 1 mL ether and 1 mL THF, wasadded diazomethane solution (generated by adding 40% potassium hydroxideaqueous solution to 2-methyl-3-nitro-nitrosoguanidine (40 mg, 0.27 mmol)in 2 mL ether at −78° C.) at 0° C., followed by 10 mg palladium (II)acetate (10 mg, 0.4 mmol). The reaction mixture was stirred at 0° C. for1 hour, then diluted with ethyl acetate and washed with brine. Theorganic layers were combined and concentrated in vacuo to give a cruderesidue, which was purified by column chromatography (silica 25-50%EtOAc/hexane) to give[2-(3-cyclopropyl-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester.

Catalytic hydrogenation of[2-(3-cyclopropyl-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester, formation of the sulfonyl urea, followed by TFAdeprotection, provided5-chloro-N-[({2-cyclopropyl-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=547, 549(Cl).

Example 95

The substituted aniline was generated by Method A (Example 46) using1-fluoro-4-nitro-2-trifluoromethyl-benzene. Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-(trifluoromethyl)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=575, 577 (Cl).

Example 96

The substituted aniline was generated by Method C (Example 48) coupling4-bromo-2,6-dimethyl-phenylamine to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester. Formation of the sulfonyl urea was achieved using themethod described in Example 10, followed by TFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2,6-dimethylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=535, 537(Cl).

Example 97

4-Fluoro-1-nitro-2-trifluoromethyl-benzene was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(trifluoromethyl)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=575, 577 (Cl).

Example 98

To a solution of 5-fluoro-2-nitro-benzoic acid (1 g, 5.4 mmol in 5 mLTHF, was added (trimethylsilyl)diazomethane (2 M in ether, 11 ml, 22mmol) at 0° C. The reaction mixture was warmed to room temperature underargon and stirred for 30 minutes, then diluted with ethyl acetate andwashed with brine. The organic layers were combined and concentrated invacuo to give a crude residue, which was purified by columnchromatography (silica 5-30% EtOAc/hexane) to give5-fluoro-2-nitro-benzoic acid methyl ester.

5-Fluoro-2-nitro-benzoic acid methyl ester was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method A (Example 46).

Formation of the sulfonyl urea was achieved using the method describedin Example 10, followed by TFA deprotection, to give methyl2-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzoate.ES-MS (M+H)⁺=565, 567(Cl).

Example 99

This analog compound was obtained from hydrolysis (as described inExample 63) of methyl2-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzoateobtained in Example 98.2-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]benzoicacid. ES-MS (M+H)⁺=551, 553(Cl).

Example 100

To a solution of 2-isopropyl-6-methyl-phenylamine (1 ml, 6.4 mmol) in6.5 mL acetic acid, was added bromine (0.33 ml, 6.4 mmol) dropwise over10 minutes at room temperature. The mixture was stirred at roomtemperature under argon for 20 minutes. A precipitate was formed andcollected by filtration and washed with ether to give pure4-bromo-2-isopropyl-6-methyl-phenyl amine.

4-Bromo-2-isopropyl-6-methyl-phenylamine was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method C (Example 48). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-isopropyl-6-methylphenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=563, 565(Cl).

Example 101

3-Methyl-2-nitro-benzoic acid methyl ester was obtained from3-methyl-2-nitro-benzoic acid using the procedure described in Example98, then reduced to 2-amino-3-methyl-benzoic acid methyl ester byhydrogenation as described in Method A (Example 46).2-Amino-5-bromo-3-methyl-benzoic acid methyl ester was obtained usingthe procedure in Example 100.

2-Amino-5-bromo-3-methyl-benzoic acid methyl ester was coupled to(7-Fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl)-methyl-carbamic acidtert-butyl ester using Method C (Example 48). Formation of the sulfonylurea was achieved using the method described in Example 10, followed byTFA deprotection, to methyl2-[({[(5-chlorothien-2-yl)sulfonyl]amino}carbonyl)amino]-5-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-3-methylbenzoate.ES-MS (M+H)⁺=579, 581 (Cl).

Example 102

[7-Fluoro-2-(3-methanesulfonyl-4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester was obtained from[2-(3-bromo-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester in Example 78.

To a slolution of[2-(3-bromo-4-nitro-phenyl)-7-fluoro-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (10 mg, 0.02 mmol) in DMSO 0.5 ml, sodiummethanesulfinate (62 mg, 0.06 mmol) was added. The reaction mixture wassubjected to microwave irradiation (temperature 12° C.) for 45 seconds.A precipitate was formed and collected by filtration to give[7-fluoro-2-(3-methanesulfonyl-4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester.

[7-fluoro-2-(3-methanesulfonyl-4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester was hydrogenated to provide an aniline. Formationof the sulfonyl urea was achieved using the method described in Example10, followed by TFA deprotection, to give5-chloro-N-({[4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]-2-(methylsulfonyl)phenyl]amino}carbonyl)thiophene-2-sulfonamide.ES-MS (M+H)⁺=585, 587(Cl).

Example 103

The substituted aniline was generated by Method C (Example 48) using2-chloro-4-iodo-phenylamine. Formation of the sulfonyl urea was achievedusing the method described in Example 10, followed by TFA deprotection,to give5-chloro-N-[({2-chloro-4-[7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=541, 543(Cl).

Example 104

[7-Fluoro-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (1.0 g, 24 mmol, from Example 8) was dissolved in12.5 mL of TFA and stirred for 0.5 h. The solvent was removed in vacuoand re-dissolved in dichloromethane/heptane and concentrated to give 1.0g (97%) of the TFA salt as a yellowish solid. ES-MS (M+H)⁺=314.2.

Example 105

To a mixture of7-Fluoro-6-methylamino-2-(4-nitro-phenyl)-2H-isoquinolin-1-one (100 mg,0.27 mmol) (Example 104) in DMF (3 mL) was added N-chlorosuccinimide (76mg, 0.57 mmol). After 18 hours all starting material was consumed. Tindichloride dehydrate 244 mg (0.11 mmol) was added to the reactionmixture, and the reaction was heated to 80° C. for 2 hr. Upon coolingthe reaction mixture was treated with 1 mL of 10% sodium carbonate andextracted 3 times with EtOAc. The combined organic layers were driedover Na₂SO₄ anhydrous and concentrated in vacuo to give a mixture of the5-Cl and 4,5-dichloro anilines. This mixture was coupled directly with76 mg (0.28 mmol) of (5-Chloro-thiophene-2-sulfonyl)-carbamic acid ethylester (Example 12) in toluene (0.54 mL) and refluxed for 4 h. Theresulting mixture of monochloro and dichloro sulfonylureas wereseparated by preparative RP-HPLC to give 9.4 mg (6% yield overall) of5-chloro-N-[({4-[5-chloro-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=541.3 (2Cl).

Example 106

The dichloro sulfonylurea was purified from the above reaction inExample 105 to afford 15 mg (10% overall yield) of5-chloro-N-[({4-[4,5-dichloro-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=576.2 (3Cl).

Example 107

7-Fluoro-6-methylamino-2-(4-nitro-phenyl)-2H-isoquinolin-1-one (800 mg,1.9 mmol, TFA salt from example 104) was partially dissolved in 18 mL ofDMF containing 517 mg (2 equiv) of K₂CO₃. RecrystallizedN-bromosuccinimide (433 mg, 2.4 mmol) was added and the reaction waswarmed to 70° C. and stirred for 18 h. The reaction mixture was quenchedwith 25 mL of water and cooled to 0° C. The resulting precipitate wascollected by filtration, washed with water, and dried in vacuo at 23° C.to give 600 mg (82%) of an off-white solid. ES-MS (M+H)⁺=392.3 (Br).

Example 108

5-Bromo-7-fluoro-6-methylamino-2-(4-nitro-phenyl)-2H-isoquinolin-1-one(400 mg, 1.0 mmol, Example 107) was combined with 920 mg (4.0 mmol) oftin dichloride dehydrate in 5 mL of DMF and heated to 70° C. for 2 h.The reaction was then cooled to 40° C. and 1 mL of 10% sodium carbonatewas added slowly along with 2 g of celite. The reaction mixture was thenextracted 3 times with EtOAc and the combined organic layers were driedover sodium sulfate, concentrated in vacuo to afford 338 mg (94%) of ayellowish solid. ES-MS (M+H)⁺=361.2 (Br).

Example 109

The sulfonylurea is prepared via the method described in Example 13.N-[({4-[5-bromo-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide. ES-MS (M+H)⁺=585.5(Cl,Br).

Example 110

To a suspension of2-(4-Amino-phenyl)-5-bromo-7-fluoro-6-methylamino-2H-isoquinolin-1-onefrom Example 108 (26 mg, 0.07 mmol) in DME (0.5 mL) was addedtetrakis(triphenylphosphine)palladium (4 mg, 5 mol %). The suspensionwas degassed and purged with Ar. K₂CO₃ (10 mg, 0.07 mmol), water (0.2mL) and the pyridine complex of 2,4,6-trivinylcyclotriboroxance (9 mg,0.04 mmol) were then added and the mixture heated to 100° C. After 30min., the reaction mixture was cooled to room temperature. The productwas then extracted with EtOAc (20 mL), washed with brine (10 mL). Thecombined organic layers was dried over sodium sulfate and concentratedin vacuo to give 26 mg of the crude product, which was used withoutadditional purification. ES-MS (M+H)⁺=310.3.

Example 111

Coupling to form the sulfonyl urea was achieved using the methoddescribed in Example 13 using Example 110 as a coupling partner to give5-chloro-N-[({4-[7-fluoro-6-(methylamino)-1-oxo-5-vinylisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=533.0, 535.0 (Cl).

Example 112

To a solution of2-(4-Amino-phenyl)-7-fluoro-6-methylamino-5-vinyl-2H-isoquinolin-1-onefrom Example 110 (27 mg, 0.09 mmol) in ethyl acetate (1 mL) and ethanol(1 mL) under Ar was added 10% Pd/C (19 mg, 0.18 mmol Pd). The mixturewas hydrogenated under 1 atm H₂ for 2 hr, filtered through Celite andconcentrated to give 26 mg of the crude product, which was used withoutadditional purification. ES-MS (M+H)⁺=312.3.

Example 113

Coupling to form the sulfonyl urea was achieved using the methoddescribed in Example 13 using2-(4-Amino-phenyl)-5-ethyl-7-fluoro-6-methylamino-2H-isoquinolin-1-onefrom previous example as a coupling partner to give5-chloro-N-[({4-[5-ethyl-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=535.0, 537.0 (Cl).

Example 114

To a suspension of2-(4-Amino-phenyl)-5-bromo-7-fluoro-6-methylamino-2H-isoquinolin-1-onefrom Example 108 (50 mg, 0.14 mmol) in toluene (0.6 mL) was addedtetrakis(triphenylphosphine)palladium (16 mg). The suspension wasdegassed and purged with Ar. K₃PO₄ (103 mg, 0.49 mmol), water (0.2 mL)and cyclopropyl boronic acid (15 mg, 0.18 mmol) were then added and themixture heated to 100° C. After reacting overnight, the mixture wascooled to r.t. The reaction mixture was then extracted with EtOAc (20mL), washed with brine (10 mL), dried over sodium sulfate andconcentrated in vacuo to give 48 mg of the crude product. The crude waspurified over silica gel to give 12 mg (25%) of the pure product. ES-MS(M+H)⁺=324.1.

Example 115

Coupling to form the sulfonyl urea was achieved using the methoddescribed in Example 13 using to give5-chloro-N-[({4-[5-cyclopropyl-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamideES-MS (M+H)⁺547.1, 549.1 (Cl).

Example 116

2-(4-Amino-phenyl)-5-bromo-7-fluoro-6-methylamino-2H-isoquinolin-1-onefrom Example 108 (50 mg, 0.14 mmol) and tributylethoxyvinyl tin (102 μL,0.28 mmol) were combined in toluene (0.7 mL) and then the mixturedegassed and purged with Ar. Tetrakis(triphenylphosphine)palladium (16mg, 10 mol %) was added and the mixture heated to 100° C. The mixturewas stirred under Ar for 3 h, after which the reaction mixture wascooled to room temperature. Water (30 mL) was added to the reactionmixture and the product extracted with EtOAc (30 mL), washed with a 5%ammonia/water solution (30 mL) and brine (30 mL), dried over sodiumsulfate and concentrated in vacuo to give 146 mg of the crudeethoxyvinyl compound. The crude mixture was then dissolved in THF (10mL), treated with 2N HCl (3 mL) and stirred at room temperature for 1.5h. The mixture was then neutralized with NaHCO₃ and extracted with EtOAc(30 mL). The EtOAc layer was washed with brine (25 mL), dried oversodium sulfate and concentrated in vacuo to give the crude ketone, whichwas purified over silica gel to give the final product. ES-MS(M+H)⁺=326.1.

Example 117

Coupling to form the sulfonyl urea was achieved using the methoddescribed in Example 13 to giveN-[({4-[5-acetyl-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]-5-chlorothiophene-2-sulfonamide.ES-MS (M+H)⁺=549.0, 551.0 (Cl).

Example 118

A 110 mg (0.28 mmol) portion of5-Bromo-7-fluoro-6-methylamino-2-(4-nitro-phenyl)-2H-isoquinolin-1-one(Example 107) and 55 mg (2.2 equiv) of CuCN in 1.4 mL of NMP was heatedto 200° C. for 2 h. After cooling to 50° C., 10 mL of 10% aq KCN wasadded, and mixture was stirred, filtered, washed with water and dried togive 123 mg (130%) of7-Fluoro-6-methylamino-2-(4-nitro-phenyl)-1-oxo-1,2-dihydro-isoquinoline-5-carbonitrile.A 50 mg (0.15 mmol) portion of this 5-cyano material was reduced with 36mg of Raney Nickel (Aldrich) in 1:1:2 water/AcOH/pyridine containing 58mg of sodium hypophosphite. The reaction was then extracted 3 times withethyl acetate, drying over Na₂SO₄ (anh.), and concentration in vacuoafforded 10 mg (20%) of the resulting aniline. This material was thencoupled using the method described in Example 13 to give 7.8 mg (46%) of5-chloro-N-[({4-[5-cyano-7-fluoro-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamide.ES-MS (M+H)⁺=532.1 (Cl).

Example 119

[7-Fluoro-1-oxo-2-(2-trimethyl-silanyl-ethoxymethyl)-1,2-dihydro-isoquinolin-6-yl]-carbamicacid tert-butyl ester (200 mg, 0.5 mmol, Example 44) was dissolved in1.5 mL of dry THF and further dried by stirring for 1 h in the presenceof 3A molecular sieves. The reaction mixture was then cooled to −78° C.and 1.0 mL of t-BuLi (1.9M) was added dropwise. After 1 h, 123 μL (4equiv) of MeI was added and the reaction mixture was warmed slowly to23° C. After addition of 2 mL of sat. NH₄Cl and extraction 3 times withethyl acetate, the combined organic layers were dried over Na₂SO₄,concentrated and purified on silica gel eluting with a 5% to 20% ethylacetate/hexane gradient to afford 44 mg (20%) of[7-Fluoro-5-methyl-1-oxo-2-(2-trimethylsilanyl-ethoxymethyl)-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester. ES-MS (M+H)⁺=437.4.

Example 120

[7-Fluoro-5-methyl-1-oxo-2-(2-trimethylsilanyl-ethoxymethyl)-1,2-dihydro-isoquinolin-6-yl]-methyl-carbamicacid tert-butyl ester (44 mg, 0.10 mmol, Example 119) was treated withTFA for 1 h and concentrated in vacuo. Treatment of this crude materialwith 25 mg (0.115 mmol) of p-iodoaniline, 4 mg of CuI, 3 mg8-hydroxyquinoline and 17 mg of K₂CO₃ in 200 μL of DMSO at 120° C. for18 hrs, after which the reaction was cooled to room temperature, dilutedwith ethyl acetate and washed with brine. The combined organic layerswere dried over Na₂SO₄ and concentrated in vacuo to give a cruderesidue, which was subjected to coupling with 57 mg of(5-Chloro-thiophene-2-sulfonyl)-carbamic acid ethyl ester as describedin Example 13, gave 10 mg of5-chloro-N-[({4-[7-fluoro-5-methyl-6-(methylamino)-1-oxoisoquinolin-2(1H)-yl]phenyl}amino)carbonyl]thiophene-2-sulfonamideafter RP-HPLC purification. ES-MS (M+H)⁺=521.1 (Cl).

Example 121

Step 1: Preparation of7-fluoro-6-(methylamino)-3,4-dihydroisoquinolin-1(2H)-one

A mixture of tert-butyl7-fluoro-1-oxo-1,2-dihydroisoquinolin-6-yl(methyl)carbamate (Example 45,700 mg, 2.40 mmol) and PtO₂ (470 mg) in MeOH (30 mL) containingconcentrated HCl (8 drops) was hydrogenated under 300 psi in a highpressure vessel overnight. The reaction mixture was then filtered andthe filtrate was concentrated in vacuo. The residue was treated with TFA(10 mL) and stirred for 1 hr, after which, the TFA solution wasconcentrated in vacuo. The residue was purified by RP-HPLC to give thenoted intermediate compound (142 mg). ES-MS (M+H)⁺=195.

Step 2: Preparation of2-(4-aminophenyl)-7-fluoro-6-(methylamino)-3,4-dihydroisoquinolin-1(2H)-one

A mixture of 7-fluoro-6-(methylamino)-3,4-dihydroisoquinolin-1(2H)-one(70 mg, 0.36 mmol), 4-iodoaniline (119 mg, 0.543 mmol), CuI (27 mg, 0.14mmol), 1,2-diaminocyclohexane (44 uL, 0.36 mmol) and K₃PO₄ (153 mg,0.722 mmol) in dioxane (1.5 mL) was heated at 110° C. overnight, thendiluted with CH₃CN (5 mL) and H₂O (5 mL) and filtered. The filtrate waspurified by RP-HPLC to give the noted intermediate compound as a solid(62 mg). ES-MS (M+H)⁺=286.

Step 3: Preparation of1-(5-chlorothiophen-2-ylsulfonyl)-3-(4-(7-fluoro-6-(methylamino)-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)phenyl)urea

To a solution of2-(4-aminophenyl)-7-fluoro-6-(methylamino)-3,4-dihydroisoquinolin-1(2H)-one(62 mg, 0.22 mmol) in HOAc (4 mL) at 100 C, ethyl5-chlorothiophen-2-ylsulfonylcarbamate (119 mg, 0.44 mmol) was added.After being stirred at room temperature for 1 h, the reaction mixturewas concentrated in vacuo. The residue was purified by RP-HPLC to givethe titled compound as a powder (25 mg). ES-MS (M+H)⁺509, 511 (Clpattern).

Example 1221-(5-chlorothiophen-2-ylsulfonyl)-3-(4-(5,7-difluoro-6-(methylamino)-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)phenyl)urea

To a solution of1-(5-chlorothiophen-2-ylsulfonyl)-3-(4-(7-fluoro-6-(methylamino)-1-oxo-3,4-dihydroisoquinolin-2(1H)-yl)phenyl)urea(17 mg, 0.033 mmol) in DMF (2 mL) at room temperature, selectFluor(1-(chloromethyl)-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate),32 mg, 0.090 mmol) was added. After being stirred at room temperaturefor 1 h, the reaction mixture was concentrated in vacuo. The residue waspurified by RP-HPLC to give the titled compound as a powder (1 mg).ES-MS (M+H)⁺=527, 529 (Cl pattern).

Example 123 This Example Provides an Assay for the Inhibition ofADP-Mediated Platelet Aggregation In Vitro Using Platelet-Rich Plasma(PRP)

The effect of compounds of the invention on ADP-induced human plateletaggregation using platelet-rich plasma (PRP) is preferably assessed in a96-well microtiter assay. Human venous blood is collected from healthy,drug-free volunteers into 0.38% sodium citrate (final concentration;e.g., 6 mL of 3.8% per 60 mL of blood). Platelet-rich plasma (PRP) isprepared by centrifugation at 160×g for 20 minutes at room temperature.PRP is collected, and the platelet concentration is determined using aCoulter counter or hemocytometer (platelet concentration should be2-4×10⁸ platelets per mL).

Inhibition of ADP-dependent aggregation is preferably determined in96-well flat-bottom microtiter plates using a microtiter plate shakerand plate reader similar to the procedure described by Frantantoni etal., Am. J. Clin. Pathol. 94:613 (1990). All steps are performed at roomtemperature. The total reaction volume of 0.2 mL/well includes: PRP(˜6×10⁷ total platelets in the presence of plasma), serial dilutions oftest compounds (buffer for control wells) in 0.6% DMSO. After about 5minutes preincubation at room temperature, ADP is added to a finalconcentration of 2 μM which induces submaximal aggregation. Buffer isadded instead of ADP to one set of control wells (ADP-control). The ODof the samples is then determined at 650 nm using a microtiter platereader (Softmax, Molecular Devices, Menlo Park, Calif.) resulting in the0 minute reading. The plates are then agitated for 5 min on a microtiterplate shaker and the 5 minute reading is obtained in the plate reader.Aggregation is calculated from the decrease of OD at 650 nm at t=5minutes compared to t=0 minutes and is expressed as % of the decrease inthe ADP control samples after correcting for changes in the unaggregatedcontrol samples.

Inhibition of [³H]2-MeS-ADP Binding to Platelets

Having first determined that the compounds according to the inventioninhibit ADP-dependent platelet aggregation with the above assay, asecond assay is used to determine whether such inhibition is mediated byinteraction with platelet ADP receptors. Utilizing the second assay thepotency of inhibition of such compounds with respect to [³H]2-MeS-ADPbinding to whole platelets is determined. [³H]2-MeS-ADP bindingexperiments are routinely performed with outdated human plateletscollected by standard procedures at hospital blood banks. Apyrase-washedoutdated platelets are prepared as follows (all steps at roomtemperature, if not indicated otherwise):

Outdated platelet suspensions are diluted with 1 volume of CGS andplatelets pelleted by centrifugation at 1900×g for 45 minutes. Plateletpellets are resuspended at 3-6×10⁹ platelets/ml in CGS containing 1 U/mlapyrase (grade V, Sigma, St. Louis, Mo.) and incubated for 15 minutes at37° C. After centrifugation at 730×g for 20 minutes, pellets areresuspended in Hepes-Tyrode's buffer containing 0.1% BSA (Sigma, St.Louis, Mo.) at a concentration of 6.66×10⁸ platelets/ml. Bindingexperiments are performed after >45 minutes resting of the platelets.

Alternatively, binding experiments are performed with fresh humanplatelets prepared as described in I. (Inhibition of ADP-MediatedPlatelet Aggregation in vitro), except that platelets are resuspended inHepes-Tyrode's buffer containing 0.1% BSA (Sigma, St. Louis, Mo.) at aconcentration of 6.66×10⁸ platelets/ml. Very similar results areobtained with fresh and outdated platelets.

A platelet ADP receptor binding assay using the tritiated potent agonistligand [³H]2-MeS-ADP (Jantzen, H. M. et al. (1999) Thromb. Hemost.81:111-117) has been adapted to the 96-well microtiter format. In anassay volume of 0.2 ml Hepes-Tyrode's buffer with 0.1% BSA and 0.6%DMSO, 1×10⁸ apyrase-washed platelets are preincubated in 96-well flatbottom microtiter plates for 5 minutes with serial dilutions of testcompounds before addition of 1 nM [³H]2-MeS-ADP([³H]2-methylthioadenosine-5′-diphosphate, ammonium salt; specificactivity 48-49 Ci/mmole, obtained by custom synthesis from Amersham LifeScience, Inc., Arlington Heights, Ill., or NEN Life Science Products,Boston, Mass.). Total binding is determined in the absence of testcompounds. Samples for nonspecific binding may contain 10⁻⁵ M unlabelled2-MeS-ADP (RBI, Natick, Mass.). After incubation for 15 minutes at roomtemperature, unbound radioligand is separated by rapid filtration andtwo washes with cold (4-8° C.) Binding Wash Buffer (10 mM Hepes pH 7.4,138 mM NaCl) using a 96-well cell harvester (Minidisc 96, SkatronInstruments, Sterling, Va.) and 8×12 GF/C glassfiber filtermats (PrintedFiltermat A, for 1450 Microbeta, Wallac Inc., Gaithersburg, Md.). Theplatelet-bound radioactivity on the filtermats is determined in ascintillation counter (Microbeta 1450, Wallac Inc., Gaithersburg, Md.).Specific binding is determined by subtraction of non-specific bindingfrom total binding, and specific binding in the presence of testcompounds is expressed as % of specific binding in the absence of testcompounds dilutions.

Example 124 This Example Provides Activity for Selected Compounds of theInvention, Evaluated as Described Above. In the Table Below, Activity inthe PRP Assay is Provided as Follows: +++, IC₅₀<10 μM; ++, 10 μM<IC₅₀<30μM; and +, IC₅₀>30 μM

Example No. Activity Example 10 +++ Example 13 ++ Example 14 + Example19 +++ Example 20 +++ Example 21 ++ Example 22 +++ Example 23 +++Example 26 + Example 27 +++ Example 28 +++ Example 29 +++ Example 30 +++Example 31 ++ Example 32 +++ Example 33 ++ Example 34 +++ Example 35 +++Example 37 +++ Example 38 +++ Example 39 ++ Example 40 + Example 41 ++Example 42 +++ Example 49 +++ Example 50 +++ Example 51 +++ Example 52+++ Example 53 +++ Example 54 +++ Example 55 +++ Example 56 +++ Example57 + Example 58 +++ Example 59 +++ Example 60 +++ Example 61 +++ Example62 ++ Example 63 +++ Example 65 ++ Example 67 ++ Example 68 ++ Example70 ++ Example 73 ++ Example 74 + Example 75 +++ Example 76 +++ Example77 +++ Example 79 + Example 80 + Example 81 +++ Example 82 ++ Example 83+++ Example 84 ++ Example 85 +++ Example 86 +++ Example 87 +++ Example88 +++ Example 89 +++ Example 90 +++ Example 91 +++ Example 92 ++Example 93 +++ Example 94 +++ Example 95 + Example 96 +++ Example 97 +++Example 98 +++ Example 99 +++ Example 100 ++ Example 101 +++ Example102 + Example 103 +++ Example 109 + Example 118 ++ Example 120 +++

It should be understood that the foregoing discussion, embodiments andexamples merely present a detailed description of certain preferredembodiments. It will be apparent to those of ordinary skill in the artthat various modifications and equivalents can be made without departingfrom the spirit and scope of the invention. All the patents, journalarticles and other documents discussed or cited above are hereinincorporated by reference.

1. A compound having the formula:

or a pharmaceutically acceptable salt thereof, wherein the dotted linerepresents an optional double bond; R¹ is a member selected from thegroup consisting of C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl and benzyl; R² is a member selected from the groupconsisting of H, C₁₋₆ alkyl and C₁₋₆ haloalkyl; R³ is a member selectedfrom the group consisting of H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₆ haloalkyl, C₁₋₆hydroxyalkyl, cyano and —C(O)R^(3a), wherein R^(3a) is a member selectedfrom the group consisting of H, hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, amino,C₁₋₆ alkylamino and di-C₁₋₆ alkylamino; R⁴ is a member selected from thegroup consisting of H and C₁₋₆ alkyl; R⁵ is a member selected from thegroup consisting of H, halogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₁₋₆ haloalkyl, C₁₋₆ alkoxy, cyano and —C(O)R^(5a), wherein R^(5a) is amember selected from the group consisting of C₁₋₆ alkoxy, amino, C₁₋₆alkylamino and di-C₁₋₆ alkylamino; and Ar is an aromatic ring selectedfrom the group consisting of benzene, pyridine and pyrimidine, each ofwhich is optionally substituted with from 1-2 R⁶ substituents, whereineach R⁶ is independently selected from the group consisting of halogen,cyano, hydroxy, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ alkoxy,C₁₋₆ haloalkyl, C₁₋₆ haloalkoxy, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl,C₃₋₅ cycloalkyl-alkoxy, amino, C₁₋₆ alkylamino, di-C₁₋₆ alkylamino,—C(O)R^(6a), —O(CH₂)_(m)OR^(6b), —(CH₂)_(m)OR^(6b),—O(CH₂)_(m)N(R^(6b))₂ and —(CH₂)_(m)N(R^(6b))₂, wherein the subscript mis an integer of from 1 to 3, each R^(6a) is a member independentlyselected from the group consisting of H, hydroxy, C₁₋₆ alkyl, C₁₋₆alkoxy, amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino, and each R^(6b)is a member independently selected from the group consisting of H, C₁₋₄alkyl and C₁₋₄ alkanoyl, and optionally, two R^(6b) groups attached tonitrogen are combined with the nitrogen atom to form an azetidine,pyrrolidine or piperidine ring.
 2. A compound of claim 1, having theformula:

wherein the subscript n is an integer of from 0 to
 2. 3. A compound ofclaim 1, having the formula:

wherein the subscript n is an integer of from 0 to
 2. 4. A compound ofclaim 1, having the formula:

wherein the subscript n is an integer of from 0 to
 2. 5. A compound ofclaim 2, wherein n is an integer of from 0 to 2; R¹ is C₁₋₄ alkyl, C₃₋₅cycloalkyl, or C₃₋₅ cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄haloalkyl, cyano or —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen,C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶,when present is selected from the group consisting of C₁₋₄ alkyl, C₁₋₄alkoxy, C₃₋₅ cycloalkyl-alkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R⁶)₂wherein the subscript m is 1 or 2 and each R^(6b) is independentlyselected from the group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.6. A compound of claim 5, wherein R¹ is C₁₋₄ alkyl; R⁴ is H or CH₃; R⁵is halogen or C₁₋₄ alkyl; and R⁶ when present is selected from C₁₋₄alkyl, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂.
 7. A compound ofclaim 6, wherein R¹ is methyl; R⁵ is chloro, and is attached at the5-position of the thienyl ring; and R⁶ when present is selected from thegroup consisting of CH₃, —OCH₂CH₂OH, —OCH₂CH₂OCH₃, —OCH₂OCH₃,—OCH₂CH₂OC(O)CH₃ and —O(CH₂)₂N(CH₃)₂.
 8. A compound of claim 7, whereinn is
 0. 9. A compound of claim 7, wherein n is
 1. 10. A compound ofclaim 7, wherein n is
 2. 11. A compound of claim 3, wherein n is 0 or 1;R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅ cycloalkyl-alkyl; R² is H; R³is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyano or —C(O)R^(3a); R⁴ is H or C₁₋₄alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄ alkoxy, C₁₋₄ haloalkyl, —CN,—C≡CH or —CONH₂; and R⁶, when present is selected from the groupconsisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅ cycloalkyl-alkoxy,—O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ wherein the subscript m is1 or 2 and each R^(6b) is independently selected from the groupconsisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 12. A compound of claim11, wherein R¹ is C₁₋₄ alkyl; R⁴ is H or CH₃; R⁵ is halogen or C₁₋₄alkyl; and R⁶ when present is selected from C₁₋₄ alkyl,—O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂.
 13. A compound of claim12, wherein R¹ is methyl; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl or C₃₋₅ cycloalkyl-alkyl; R⁴ is H or CH₃; R⁵ ischloro and is attached at the 5-position of the thienyl ring; and R⁶,when present is selected from the group consisting of C₁₋₄ alkyl,—O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ wherein the subscript m is1 or 2 and each R^(6b) is independently selected from the groupconsisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 14. A compound of claim4, wherein n is 0 or 1; R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoor —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkylalkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ whereinthe subscript m is 1 or 2 and each R^(6b) is independently selected fromthe group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 15. A compoundof claim 14, wherein R¹ is C₁₋₄ alkyl; R⁴ is H or CH₃; R⁵ is halogen orC₁₋₄ alkyl; and R⁶ when present is selected from C₁₋₄ alkyl,—O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂.
 16. A compound of claim15, wherein R¹ is methyl; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl or C₃₋₅ cycloalkyl-alkyl; R⁴ is H or CH₃; R⁵ ischloro and is attached at the 5-position of the thienyl ring; and R⁶,when present is selected from the group consisting of C₁₋₄ alkyl,—O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ wherein the subscript m is1 or 2 and each R^(6b) is independently selected from the groupconsisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 17. A compound of claim1, selected from the group consisting of:


18. A pharmaceutical composition comprising a pharmaceuticallyacceptable excipient and a compound having the formula:

or a pharmaceutically acceptable salt thereof, wherein the dotted linerepresents an optional double bond; R¹ is a member selected from thegroup consisting of C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl and benzyl; R² is a member selected from the groupconsisting of H, C₁₋₆ alkyl and C₁₋₆ haloalkyl; R³ is a member selectedfrom the group consisting of H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₆ haloalkyl, C₁₋₆hydroxyalkyl, cyano and —C(O)R^(3a), wherein R^(3a) is a member selectedfrom the group consisting of H, hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, amino,C₁₋₆ alkylamino and di-C₁₋₆ alkylamino; R⁴ is a member selected from thegroup consisting of H and C₁₋₆ alkyl; R⁵ is a member selected from thegroup consisting of H, halogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C-6 haloalkyl, C₁₋₆ alkoxy, cyano and —C(O)R^(5a), wherein R^(5a) is amember selected from the group consisting of C₁₋₆ alkoxy, amino, C₁₋₆alkylamino and di-C-6 alkylamino; and Ar is an aromatic ring selectedfrom the group consisting of benzene, pyridine and pyrimidine, each ofwhich is optionally substituted with from 1-2 R⁶ substituents, whereineach R⁶ is independently selected from the group consisting of halogen,cyano, hydroxy, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ alkoxy,C₁₋₆ haloalkyl, C₁₋₆ haloalkoxy, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl,C₃₋₅ cycloalkyl-alkoxy, amino, C₁₋₆ alkylamino, di-C₁₋₆ alkylamino,—C(O)R^(6a), —O(CH₂)_(m)OR^(6b), —(CH₂)_(m)OR^(6b),—O(CH₂)_(m)N(R^(6b))₂ and —(CH₂)_(m)N(R^(6b))₂, wherein the subscript mis an integer of from 1 to 3, each R^(6a) is a member independentlyselected from the group consisting of H, hydroxy, C₁₋₆ alkyl, C₁₋₆alkoxy, amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino, and each R^(6b)is a member independently selected from the group consisting of H, C₁₋₄alkyl and C₁₋₄ alkanoyl, and optionally, two R^(6b) groups attached tonitrogen are combined with the nitrogen atom to form an azetidine,pyrrolidine or piperidine ring.
 19. A pharmaceutical composition ofclaim 18, wherein said compound has the formula:

wherein n is 0 or 1; R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoand —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkyl-alkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ whereinthe subscript m is 1 or 2 and each R^(6b) is independently selected fromthe group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 20. Apharmaceutical composition of claim 19, wherein R¹ is methyl; R³ is H,C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl or C₃₋₅cycloalkyl-alkyl; R⁴ is H or CH₃; R⁵ is chloro and is attached at the5-position of the thienyl ring; and R⁶, when present is selected fromthe group consisting of C₁₋₄ alkyl, —O(CH₂)_(m)OR^(6b) and—O(CH₂)_(m)N(R^(6b))₂ wherein the subscript m is 1 or 2 and each R^(6b)is independently selected from the group consisting of H, C₁₋₄ alkyl andC₁₋₄ alkanoyl.
 21. A pharmaceutical composition of claim 18, whereinsaid compound is selected from the group consisting of:


22. A method of treating thrombosis in a subject comprisingadministering to a subject in need thereof, a therapeutically effectiveamount of a compound having the formula:

or a pharmaceutically acceptable salt thereof, wherein the dotted lineindicates an optional double bond; R¹ is a member selected from thegroup consisting of C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₃₋₅ cycloalkyl, C₃₋₅cycloalkyl-alkyl and benzyl: R² is a member selected from the groupconsisting of H, C₁₋₆ alkyl and C₁₋₆ haloalkyl; R³ is a member selectedfrom the group consisting of H, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₃₋₅ cycloalkyl-alkyl, C₁₋₆ haloalkyl, C₁₋₆ hydroxyalkyl, cyano and—C(O)R^(3a), wherein R^(3a) is a member selected from the groupconsisting of H, hydroxy, C₁₋₆ alkyl, C₁₋₆ alkoxy, amino, C₁₋₆alkylamino and di-C₁₋₆ alkylamino; R⁴ is a member selected from thegroup consisting of H and C₁₋₆ alkyl; R⁵ is a member selected from thegroup consisting of H, halogen, C₁₋₆, alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,C₁₋₆ haloalkyl, C₁₋₆ alkoxy, cyano and —C(O)R^(5a), wherein R^(5a) is amember selected from the group consisting of C₁₋₆ alkoxy, amino, C₁₋₆alkylamino and di-C₁₋₆ alkylamino; and Ar is an aromatic ring selectedfrom the group consisting of benzene, pyridine and pyrimidine, each ofwhich is optionally substituted with from 1-2 R⁶ substituents, whereineach R⁶ is independently selected from the group consisting of halogen,cyano, hydroxy, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₁₋₆ alkoxy,C₁₋₆ haloalkyl, C₁₋₆ haloalkoxy, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl,C₃₋₅ cycloalkyl-alkoxy, amino, C₁₋₆ alkylamino, di-C₁₋₆ alkylamino,—C(O)R^(6a), —O(CH₂)_(m)OR^(6b), —(CH₂)_(m)OR^(6b),—O(CH₂)_(m)N(R^(6b))₂ and —(CH₂)_(m)N(R^(6b))₂, wherein the subscript mis an integer of from 1 to 3, each R^(6a) is a member independentlyselected from the group consisting of H, hydroxy, C₁₋₆ alkyl, C₁₋₆alkoxy, amino, C₁₋₆ alkylamino and di-C₁₋₆ alkylamino, and each R^(6b)is a member independently selected from the group consisting of H, C₁₋₄alkyl and C₁₋₄ alkanoyl, and optionally, two R^(6b) groups attached tonitrogen are combined with the nitrogen atom to form an azetidine,pyrrolidine or piperidine ring.
 23. A method in accordance with claim22, wherein said compound is administered in combination with a secondtherapeutic agent selected from the group consisting of antiplateletcompounds, anticoagulants, fibrinolytics, anti-inflammatory compounds,cholesterol-lowering agents, blood pressure-lowering agents andserotonin blockers.
 24. A method in accordance with claim 23, whereinsaid second therapeutic agent is an antiplatelet compound selected fromthe group consisting of GPIIB-IIIa antagonists, aspirin,phosphodiesterase III inhibitors and thromboxane A2 receptorantoagonists.
 25. A method in accordance with claim 23, wherein saidsecond therapeutic agent is an anticoagulant selected from the groupconsisting of thrombin inhibitors, coumadin, heparin and Lovenox®.
 26. Amethod in accordance with claim 23, wherein said second therapeuticagent is an anti-inflammatory compound selected from the groupconsisting of non-steroidal anti-inflammatory agents, cyclooxygenase-2inhibitors and rheumatoid arthritis agents.
 27. A method in accordancewith claim 23, wherein said compound is administered orally.
 28. Amethod for preventing the occurrence of a secondary ischemic eventcomprising administering to a patient who has suffered a primaryischemic event a therapeutically effective amount of a compound of claim1, together with a pharmaceutically acceptable carrier.
 29. A method inaccordance with claim 28, wherein said primary and/or secondary ischemicevent is selected from the group consisting of myocardial infarction,stable or unstable angina, acute reocclusion after percutaneoustransluminal coronary angioplasty, restenosis, thrombotic stroke,transient ischemic attack, reversible ischemic neurological deficit andintermittent claudication.
 30. A method in accordance with claim 22,wherein said compound has the formula:

wherein n is 0 or 1; R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoor —C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₁₋₄alkoxy, C₁₋₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkyl-alkoxy, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ whereinthe subscript m is 1 or 2 and each R^(6b) is independently selected fromthe group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 31. A method inaccordance with claim 30, wherein R¹ is methyl; R³ is H, C₁₋₄ alkyl,C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl or C₃₋₅ cycloalkyl-alkyl; R⁴is H or CH₃; R⁵ is chloro and is attached at the 5-position of thethienyl ring; and R⁶, when present is selected from the group consistingof C₁₋₄ alkyl, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ wherein thesubscript m is 1 or 2 and each R^(6b) is independently selected from thegroup consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 32. A method inaccordance with claim 22, wherein said compound is selected from thegroup consisting of:


33. A method in accordance with claim 28, wherein said compound has theformula:

wherein n is 0 or 1, R¹ is C₁₋₄ alkyl, C₃₋₅ cycloalkyl, or C₃₋₅cycloalkyl-alkyl; R² is H; R³ is H, C₁₋₄ alkyl, C₂₋₄ alkenyl, C₂₋₄alkynyl, C₃₋₅ cycloalkyl, C₃₋₅ cycloalkyl-alkyl, C₁₋₄ haloalkyl, cyanoor C(O)R^(3a); R⁴ is H or C₁₋₄ alkyl; R⁵ is halogen, C₁₋₄ alkyl, C₄alkoxy, C₄ haloalkyl, —CN, —C≡CH or —CONH₂; and R⁶, when present isselected from the group consisting of C₁₋₄ alkyl, C₁₋₄ alkoxy, C₃₋₅cycloalkyl-alkoxy, O(CH₂)_(m)OR^(6b) and O(CH₂)_(m)N(R^(6b))₂ whereinthe subscript m is 1 or 2 and each R^(6b) is independently selected fromthe group consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 34. A method inaccordance with claim 33, wherein R¹ is methyl; R³ is H, C₁₋₄ alkyl,C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₃₋₅ cycloalkyl or C₃₋₅ cycloalkyl-alkyl; R⁴is H or CH₃; R⁵ is chloro and is attached at the 5-position of thethienyl ring; and R⁶, when present is selected from the group consistingof C₁₋₄ alkyl, —O(CH₂)_(m)OR^(6b) and —O(CH₂)_(m)N(R^(6b))₂ wherein thesubscript m is 1 or 2 and each R^(6b) is independently selected from thegroup consisting of H, C₁₋₄ alkyl and C₁₋₄ alkanoyl.
 35. A method inaccordance with claim 28, wherein said compound is selected from thegroup consisting of: